                         SEQUENCE LISTING

<110>  Merz Pharma GmbH & Co.KGaA
 
<120>  Method for the determination of Botulinum neurotoxin biological 
       activity

<130>  MP10047PC

<160>  16    

<170>  PatentIn version 3.3

<210>  1
<211>  255
<212>  PRT
<213>  artificial

<220>
<223>  caspase mutant

<400>  1

Met Asn Ser Tyr Lys Met Asp Tyr Pro Glu Met Gly Leu Cys Ile Ile 
1               5                   10                  15      


Ile Asn Asn Lys Asn Phe His Lys Ser Thr Gly Met Thr Ser Arg Ser 
            20                  25                  30          


Gly Thr Asp Val Asp Ala Ala Asn Leu Arg Glu Thr Phe Arg Asn Leu 
        35                  40                  45              


Lys Tyr Glu Val Arg Asn Lys Asn Asp Leu Thr Arg Glu Glu Ile Val 
    50                  55                  60                  


Glu Leu Met Arg Asp Val Ser Lys Glu Asp His Ser Lys Arg Ser Ser 
65                  70                  75                  80  


Phe Val Cys Val Leu Leu Ser His Gly Glu Glu Gly Ile Ile Phe Gly 
                85                  90                  95      


Thr Asn Gly Pro Val Asp Leu Lys Lys Ile Thr Asn Phe Phe Arg Gly 
            100                 105                 110         


Asp Arg Cys Arg Ser Leu Thr Gly Lys Pro Lys Leu Phe Ile Ile Gln 
        115                 120                 125             


Ala Cys Arg Gly Thr Glu Leu Asp Cys Gly Ile Glu Gly Gly Gly Thr 
    130                 135                 140                 


Arg Ile Asp Glu Ala Asn Gln Arg Ala Thr Lys Met Leu Gly Ser Gly 
145                 150                 155                 160 


Gly Gly His Lys Ile Pro Val Glu Ala Asp Phe Leu Tyr Ala Tyr Ser 
                165                 170                 175     


Thr Ala Pro Gly Tyr Tyr Ser Trp Arg Asn Ser Lys Asp Gly Ser Trp 
            180                 185                 190         


Phe Ile Gln Ser Leu Cys Ala Met Leu Lys Gln Tyr Ala Asp Lys Leu 
        195                 200                 205             


Glu Phe Met His Ile Leu Thr Arg Val Asn Arg Lys Val Ala Thr Glu 
    210                 215                 220                 


Phe Glu Ser Phe Ser Phe Asp Ala Thr Phe His Ala Lys Lys Gln Ile 
225                 230                 235                 240 


Pro Cys Ile Val Ser Met Leu Thr Lys Glu Leu Tyr Phe Tyr His 
                245                 250                 255 


<210>  2
<211>  252
<212>  PRT
<213>  artificial

<220>
<223>  caspase mutant

<400>  2

Met Asn Ser Tyr Lys Met Asp Tyr Pro Glu Met Gly Leu Cys Ile Ile 
1               5                   10                  15      


Ile Asn Asn Lys Asn Phe His Lys Ser Thr Gly Met Thr Ser Arg Ser 
            20                  25                  30          


Gly Thr Asp Val Asp Ala Ala Asn Leu Arg Glu Thr Phe Arg Asn Leu 
        35                  40                  45              


Lys Tyr Glu Val Arg Asn Lys Asn Asp Leu Thr Arg Glu Glu Ile Val 
    50                  55                  60                  


Glu Leu Met Arg Asp Val Ser Lys Glu Asp His Ser Lys Arg Ser Ser 
65                  70                  75                  80  


Phe Val Cys Val Leu Leu Ser His Gly Glu Glu Gly Ile Ile Phe Gly 
                85                  90                  95      


Thr Asn Gly Pro Val Asp Leu Lys Lys Ile Thr Asn Phe Phe Arg Gly 
            100                 105                 110         


Asp Arg Cys Arg Ser Leu Thr Gly Lys Pro Lys Leu Phe Ile Ile Gln 
        115                 120                 125             


Ala Cys Arg Gly Thr Glu Leu Asp Cys Gly Ile Glu Gly Gly Gly Gly 
    130                 135                 140                 


Ile Asp Glu Ala Asn Gln Arg Ala Thr Lys Met Gly Gly Gly Gly His 
145                 150                 155                 160 


Lys Ile Pro Val Glu Ala Asp Phe Leu Tyr Ala Tyr Ser Thr Ala Pro 
                165                 170                 175     


Gly Tyr Tyr Ser Trp Arg Asn Ser Lys Asp Gly Ser Trp Phe Ile Gln 
            180                 185                 190         


Ser Leu Cys Ala Met Leu Lys Gln Tyr Ala Asp Lys Leu Glu Phe Met 
        195                 200                 205             


His Ile Leu Thr Arg Val Asn Arg Lys Val Ala Thr Glu Phe Glu Ser 
    210                 215                 220                 


Phe Ser Phe Asp Ala Thr Phe His Ala Lys Lys Gln Ile Pro Cys Ile 
225                 230                 235                 240 


Val Ser Met Leu Thr Lys Glu Leu Tyr Phe Tyr His 
                245                 250         


<210>  3
<211>  171
<212>  PRT
<213>  artificial

<220>
<223>  caspase mutant

<400>  3

Met Asn Ser Tyr Lys Met Asp Tyr Pro Glu Met Gly Leu Cys Ile Ile 
1               5                   10                  15      


Ile Asn Asn Lys Asn Phe His Lys Ser Thr Gly Met Thr Ser Arg Ser 
            20                  25                  30          


Gly Thr Asp Val Asp Ala Ala Asn Leu Arg Glu Thr Phe Arg Asn Leu 
        35                  40                  45              


Lys Tyr Glu Val Arg Asn Lys Asn Asp Leu Thr Arg Glu Glu Ile Val 
    50                  55                  60                  


Glu Leu Met Arg Asp Val Ser Lys Glu Asp His Ser Lys Arg Ser Ser 
65                  70                  75                  80  


Phe Val Cys Val Leu Leu Ser His Gly Glu Glu Gly Ile Ile Phe Gly 
                85                  90                  95      


Thr Asn Gly Pro Val Asp Leu Lys Lys Ile Thr Asn Phe Phe Arg Gly 
            100                 105                 110         


Asp Arg Cys Arg Ser Leu Thr Gly Lys Pro Lys Leu Phe Ile Ile Gln 
        115                 120                 125             


Ala Cys Arg Gly Thr Glu Leu Asp Cys Gly Ile Glu Ile Asp Glu Ala 
    130                 135                 140                 


Asn Gln Arg Ala Thr Lys Met His Lys Ile Pro Val Glu Ala Asp Phe 
145                 150                 155                 160 


Leu Tyr Ala Tyr Ser Thr Ala Pro Gly Tyr His 
                165                 170     


<210>  4
<211>  292
<212>  PRT
<213>  artificial

<220>
<223>  caspase mutant

<400>  4

Met Glu Asn Thr Glu Asn Ser Val Asp Ser Lys Ser Ile Lys Asn Leu 
1               5                   10                  15      


Glu Pro Lys Ile Ile His Gly Ser Glu Ser Met Asp Ser Gly Ile Ser 
            20                  25                  30          


Leu Asp Asn Ser Tyr Lys Met Asp Tyr Pro Glu Met Gly Leu Cys Ile 
        35                  40                  45              


Ile Ile Asn Asn Lys Asn Phe His Lys Ser Thr Gly Met Thr Ser Arg 
    50                  55                  60                  


Ser Gly Thr Asp Val Asp Ala Ala Asn Leu Arg Glu Thr Phe Arg Asn 
65                  70                  75                  80  


Leu Lys Tyr Glu Val Arg Asn Lys Asn Asp Leu Thr Arg Glu Glu Ile 
                85                  90                  95      


Val Glu Leu Met Arg Asp Val Ser Lys Glu Asp His Ser Lys Arg Ser 
            100                 105                 110         


Ser Phe Val Cys Val Leu Leu Ser His Gly Glu Glu Gly Ile Ile Phe 
        115                 120                 125             


Gly Thr Asn Gly Pro Val Asp Leu Lys Lys Ile Thr Asn Phe Phe Arg 
    130                 135                 140                 


Gly Asp Arg Cys Arg Ser Leu Thr Gly Lys Pro Lys Leu Phe Ile Ile 
145                 150                 155                 160 


Gln Ala Cys Arg Gly Thr Glu Leu Asp Cys Gly Ile Glu Thr Asp Ser 
                165                 170                 175     


Gly Ile Asp Glu Ala Asn Gln Arg Ala Thr Lys Met Leu Gly Ser Gly 
            180                 185                 190         


Val Asp Asp Asp Met Ala Cys His Lys Ile Pro Val Glu Ala Asp Phe 
        195                 200                 205             


Leu Tyr Ala Tyr Ser Thr Ala Pro Gly Tyr Tyr Ser Trp Arg Asn Ser 
    210                 215                 220                 


Lys Asp Gly Ser Trp Phe Ile Gln Ser Leu Cys Ala Met Leu Lys Gln 
225                 230                 235                 240 


Tyr Ala Asp Lys Leu Glu Phe Met His Ile Leu Thr Arg Val Asn Arg 
                245                 250                 255     


Lys Val Ala Thr Glu Phe Glu Ser Phe Ser Phe Asp Ala Thr Phe His 
            260                 265                 270         


Ala Lys Lys Gln Ile Pro Cys Ile Val Ser Met Leu Thr Lys Glu Leu 
        275                 280                 285             


Tyr Phe Tyr His 
    290         


<210>  5
<211>  288
<212>  PRT
<213>  artificial

<220>
<223>  caspase mutant

<400>  5

Met Glu Asn Thr Glu Asn Ser Val Asp Ser Lys Ser Ile Lys Asn Leu 
1               5                   10                  15      


Glu Pro Lys Ile Ile His Gly Ser Glu Ser Met Asp Ser Gly Ile Ser 
            20                  25                  30          


Leu Asp Asn Ser Tyr Lys Met Asp Tyr Pro Glu Met Gly Leu Cys Ile 
        35                  40                  45              


Ile Ile Asn Asn Lys Asn Phe His Lys Ser Thr Gly Met Thr Ser Arg 
    50                  55                  60                  


Ser Gly Thr Asp Val Asp Ala Ala Asn Leu Arg Glu Thr Phe Arg Asn 
65                  70                  75                  80  


Leu Lys Tyr Glu Val Arg Asn Lys Asn Asp Leu Thr Arg Glu Glu Ile 
                85                  90                  95      


Val Glu Leu Met Arg Asp Val Ser Lys Glu Asp His Ser Lys Arg Ser 
            100                 105                 110         


Ser Phe Val Cys Val Leu Leu Ser His Gly Glu Glu Gly Ile Ile Phe 
        115                 120                 125             


Gly Thr Asn Gly Pro Val Asp Leu Lys Lys Ile Thr Asn Phe Phe Arg 
    130                 135                 140                 


Gly Asp Arg Cys Arg Ser Leu Thr Gly Lys Pro Lys Leu Phe Ile Ile 
145                 150                 155                 160 


Gln Ala Cys Arg Gly Thr Glu Leu Asp Cys Gly Ile Glu Thr Asp Ser 
                165                 170                 175     


Gly Ile Asp Glu Ala Asn Gln Arg Ala Thr Lys Met Val Asp Asp Asp 
            180                 185                 190         


Met Ala Cys His Lys Ile Pro Val Glu Ala Asp Phe Leu Tyr Ala Tyr 
        195                 200                 205             


Ser Thr Ala Pro Gly Tyr Tyr Ser Trp Arg Asn Ser Lys Asp Gly Ser 
    210                 215                 220                 


Trp Phe Ile Gln Ser Leu Cys Ala Met Leu Lys Gln Tyr Ala Asp Lys 
225                 230                 235                 240 


Leu Glu Phe Met His Ile Leu Thr Arg Val Asn Arg Lys Val Ala Thr 
                245                 250                 255     


Glu Phe Glu Ser Phe Ser Phe Asp Ala Thr Phe His Ala Lys Lys Gln 
            260                 265                 270         


Ile Pro Cys Ile Val Ser Met Leu Thr Lys Glu Leu Tyr Phe Tyr His 
        275                 280                 285             


<210>  6
<211>  416
<212>  PRT
<213>  Homo sapiens

<400>  6

Met Asp Glu Ala Asp Arg Arg Leu Leu Arg Arg Cys Arg Leu Arg Leu 
1               5                   10                  15      


Val Glu Glu Leu Gln Val Asp Gln Leu Trp Asp Ala Leu Leu Ser Arg 
            20                  25                  30          


Glu Leu Phe Arg Pro His Met Ile Glu Asp Ile Gln Arg Ala Gly Ser 
        35                  40                  45              


Gly Ser Arg Arg Asp Gln Ala Arg Gln Leu Ile Ile Asp Leu Glu Thr 
    50                  55                  60                  


Arg Gly Ser Gln Ala Leu Pro Leu Phe Ile Ser Cys Leu Glu Asp Thr 
65                  70                  75                  80  


Gly Gln Asp Met Leu Ala Ser Phe Leu Arg Thr Asn Arg Gln Ala Ala 
                85                  90                  95      


Lys Leu Ser Lys Pro Thr Leu Glu Asn Leu Thr Pro Val Val Leu Arg 
            100                 105                 110         


Pro Glu Ile Arg Lys Pro Glu Val Leu Arg Pro Glu Thr Pro Arg Pro 
        115                 120                 125             


Val Asp Ile Gly Ser Gly Gly Phe Gly Asp Val Gly Ala Leu Glu Ser 
    130                 135                 140                 


Leu Arg Gly Asn Ala Asp Leu Ala Tyr Ile Leu Ser Met Glu Pro Cys 
145                 150                 155                 160 


Gly His Cys Leu Ile Ile Asn Asn Val Asn Phe Cys Arg Glu Ser Gly 
                165                 170                 175     


Leu Arg Thr Arg Thr Gly Ser Asn Ile Asp Cys Glu Lys Leu Arg Arg 
            180                 185                 190         


Arg Phe Ser Ser Leu His Phe Met Val Glu Val Lys Gly Asp Leu Thr 
        195                 200                 205             


Ala Lys Lys Met Val Leu Ala Leu Leu Glu Leu Ala Gln Gln Asp His 
    210                 215                 220                 


Gly Ala Leu Asp Cys Cys Val Val Val Ile Leu Ser His Gly Cys Gln 
225                 230                 235                 240 


Ala Ser His Leu Gln Phe Pro Gly Ala Val Tyr Gly Thr Asp Gly Cys 
                245                 250                 255     


Pro Val Ser Val Glu Lys Ile Val Asn Ile Phe Asn Gly Thr Ser Cys 
            260                 265                 270         


Pro Ser Leu Gly Gly Lys Pro Lys Leu Phe Phe Ile Gln Ala Cys Gly 
        275                 280                 285             


Gly Glu Gln Lys Asp His Gly Phe Glu Val Ala Ser Thr Ser Pro Glu 
    290                 295                 300                 


Asp Glu Ser Pro Gly Ser Asn Pro Glu Pro Asp Ala Thr Pro Phe Gln 
305                 310                 315                 320 


Glu Gly Leu Arg Thr Phe Asp Gln Leu Asp Ala Ile Ser Ser Leu Pro 
                325                 330                 335     


Thr Pro Ser Asp Ile Phe Val Ser Tyr Ser Thr Phe Pro Gly Phe Val 
            340                 345                 350         


Ser Trp Arg Asp Pro Lys Ser Gly Ser Trp Tyr Val Glu Thr Leu Asp 
        355                 360                 365             


Asp Ile Phe Glu Gln Trp Ala His Ser Glu Asp Leu Gln Ser Leu Leu 
    370                 375                 380                 


Leu Arg Val Ala Asn Ala Val Ser Val Lys Gly Ile Tyr Lys Gln Met 
385                 390                 395                 400 


Pro Gly Cys Phe Asn Phe Leu Arg Lys Lys Leu Phe Phe Lys Thr Ser 
                405                 410                 415     


<210>  7
<211>  277
<212>  PRT
<213>  Homo sapiens

<400>  7

Met Glu Asn Thr Glu Asn Ser Val Asp Ser Lys Ser Ile Lys Asn Leu 
1               5                   10                  15      


Glu Pro Lys Ile Ile His Gly Ser Glu Ser Met Asp Ser Gly Ile Ser 
            20                  25                  30          


Leu Asp Asn Ser Tyr Lys Met Asp Tyr Pro Glu Met Gly Leu Cys Ile 
        35                  40                  45              


Ile Ile Asn Asn Lys Asn Phe His Lys Ser Thr Gly Met Thr Ser Arg 
    50                  55                  60                  


Ser Gly Thr Asp Val Asp Ala Ala Asn Leu Arg Glu Thr Phe Arg Asn 
65                  70                  75                  80  


Leu Lys Tyr Glu Val Arg Asn Lys Asn Asp Leu Thr Arg Glu Glu Ile 
                85                  90                  95      


Val Glu Leu Met Arg Asp Val Ser Lys Glu Asp His Ser Lys Arg Ser 
            100                 105                 110         


Ser Phe Val Cys Val Leu Leu Ser His Gly Glu Glu Gly Ile Ile Phe 
        115                 120                 125             


Gly Thr Asn Gly Pro Val Asp Leu Lys Lys Ile Thr Asn Phe Phe Arg 
    130                 135                 140                 


Gly Asp Arg Cys Arg Ser Leu Thr Gly Lys Pro Lys Leu Phe Ile Ile 
145                 150                 155                 160 


Gln Ala Cys Arg Gly Thr Glu Leu Asp Cys Gly Ile Glu Thr Asp Ser 
                165                 170                 175     


Gly Val Asp Asp Asp Met Ala Cys His Lys Ile Pro Val Glu Ala Asp 
            180                 185                 190         


Phe Leu Tyr Ala Tyr Ser Thr Ala Pro Gly Tyr Tyr Ser Trp Arg Asn 
        195                 200                 205             


Ser Lys Asp Gly Ser Trp Phe Ile Gln Ser Leu Cys Ala Met Leu Lys 
    210                 215                 220                 


Gln Tyr Ala Asp Lys Leu Glu Phe Met His Ile Leu Thr Arg Val Asn 
225                 230                 235                 240 


Arg Lys Val Ala Thr Glu Phe Glu Ser Phe Ser Phe Asp Ala Thr Phe 
                245                 250                 255     


His Ala Lys Lys Gln Ile Pro Cys Ile Val Ser Met Leu Thr Lys Glu 
            260                 265                 270         


Leu Tyr Phe Tyr His 
        275         


<210>  8
<211>  479
<212>  PRT
<213>  Homo sapiens

<400>  8

Met Asp Phe Ser Arg Asn Leu Tyr Asp Ile Gly Glu Gln Leu Asp Ser 
1               5                   10                  15      


Glu Asp Leu Ala Ser Leu Lys Phe Leu Ser Leu Asp Tyr Ile Pro Gln 
            20                  25                  30          


Arg Lys Gln Glu Pro Ile Lys Asp Ala Leu Met Leu Phe Gln Arg Leu 
        35                  40                  45              


Gln Glu Lys Arg Met Leu Glu Glu Ser Asn Leu Ser Phe Leu Lys Glu 
    50                  55                  60                  


Leu Leu Phe Arg Ile Asn Arg Leu Asp Leu Leu Ile Thr Tyr Leu Asn 
65                  70                  75                  80  


Thr Arg Lys Glu Glu Met Glu Arg Glu Leu Gln Thr Pro Gly Arg Ala 
                85                  90                  95      


Gln Ile Ser Ala Tyr Arg Val Met Leu Tyr Gln Ile Ser Glu Glu Val 
            100                 105                 110         


Ser Arg Ser Glu Leu Arg Ser Phe Lys Phe Leu Leu Gln Glu Glu Ile 
        115                 120                 125             


Ser Lys Cys Lys Leu Asp Asp Asp Met Asn Leu Leu Asp Ile Phe Ile 
    130                 135                 140                 


Glu Met Glu Lys Arg Val Ile Leu Gly Glu Gly Lys Leu Asp Ile Leu 
145                 150                 155                 160 


Lys Arg Val Cys Ala Gln Ile Asn Lys Ser Leu Leu Lys Ile Ile Asn 
                165                 170                 175     


Asp Tyr Glu Glu Phe Ser Lys Glu Arg Ser Ser Ser Leu Glu Gly Ser 
            180                 185                 190         


Pro Asp Glu Phe Ser Asn Gly Glu Glu Leu Cys Gly Val Met Thr Ile 
        195                 200                 205             


Ser Asp Ser Pro Arg Glu Gln Asp Ser Glu Ser Gln Thr Leu Asp Lys 
    210                 215                 220                 


Val Tyr Gln Met Lys Ser Lys Pro Arg Gly Tyr Cys Leu Ile Ile Asn 
225                 230                 235                 240 


Asn His Asn Phe Ala Lys Ala Arg Glu Lys Val Pro Lys Leu His Ser 
                245                 250                 255     


Ile Arg Asp Arg Asn Gly Thr His Leu Asp Ala Gly Ala Leu Thr Thr 
            260                 265                 270         


Thr Phe Glu Glu Leu His Phe Glu Ile Lys Pro His Asp Asp Cys Thr 
        275                 280                 285             


Val Glu Gln Ile Tyr Glu Ile Leu Lys Ile Tyr Gln Leu Met Asp His 
    290                 295                 300                 


Ser Asn Met Asp Cys Phe Ile Cys Cys Ile Leu Ser His Gly Asp Lys 
305                 310                 315                 320 


Gly Ile Ile Tyr Gly Thr Asp Gly Gln Glu Ala Pro Ile Tyr Glu Leu 
                325                 330                 335     


Thr Ser Gln Phe Thr Gly Leu Lys Cys Pro Ser Leu Ala Gly Lys Pro 
            340                 345                 350         


Lys Val Phe Phe Ile Gln Ala Cys Gln Gly Asp Asn Tyr Gln Lys Gly 
        355                 360                 365             


Ile Pro Val Glu Thr Asp Ser Glu Glu Gln Pro Tyr Leu Glu Met Asp 
    370                 375                 380                 


Leu Ser Ser Pro Gln Thr Arg Tyr Ile Pro Asp Glu Ala Asp Phe Leu 
385                 390                 395                 400 


Leu Gly Met Ala Thr Val Asn Asn Cys Val Ser Tyr Arg Asn Pro Ala 
                405                 410                 415     


Glu Gly Thr Trp Tyr Ile Gln Ser Leu Cys Gln Ser Leu Arg Glu Arg 
            420                 425                 430         


Cys Pro Arg Gly Asp Asp Ile Leu Thr Ile Leu Thr Glu Val Asn Tyr 
        435                 440                 445             


Glu Val Ser Asn Lys Asp Asp Lys Lys Asn Met Gly Lys Gln Met Pro 
    450                 455                 460                 


Gln Pro Thr Phe Thr Leu Arg Lys Lys Leu Val Phe Pro Ser Asp 
465                 470                 475                 


<210>  9
<211>  521
<212>  PRT
<213>  Homo sapiens

<400>  9

Met Lys Ser Gln Gly Gln His Trp Tyr Ser Ser Ser Asp Lys Asn Cys 
1               5                   10                  15      


Lys Val Ser Phe Arg Glu Lys Leu Leu Ile Ile Asp Ser Asn Leu Gly 
            20                  25                  30          


Val Gln Asp Val Glu Asn Leu Lys Phe Leu Cys Ile Gly Leu Val Pro 
        35                  40                  45              


Asn Lys Lys Leu Glu Lys Ser Ser Ser Ala Ser Asp Val Phe Glu His 
    50                  55                  60                  


Leu Leu Ala Glu Asp Leu Leu Ser Glu Glu Asp Pro Phe Phe Leu Ala 
65                  70                  75                  80  


Glu Leu Leu Tyr Ile Ile Arg Gln Lys Lys Leu Leu Gln His Leu Asn 
                85                  90                  95      


Cys Thr Lys Glu Glu Val Glu Arg Leu Leu Pro Thr Arg Gln Arg Val 
            100                 105                 110         


Ser Leu Phe Arg Asn Leu Leu Tyr Glu Leu Ser Glu Gly Ile Asp Ser 
        115                 120                 125             


Glu Asn Leu Lys Asp Met Ile Phe Leu Leu Lys Asp Ser Leu Pro Lys 
    130                 135                 140                 


Thr Glu Met Thr Ser Leu Ser Phe Leu Ala Phe Leu Glu Lys Gln Gly 
145                 150                 155                 160 


Lys Ile Asp Glu Asp Asn Leu Thr Cys Leu Glu Asp Leu Cys Lys Thr 
                165                 170                 175     


Val Val Pro Lys Leu Leu Arg Asn Ile Glu Lys Tyr Lys Arg Glu Lys 
            180                 185                 190         


Ala Ile Gln Ile Val Thr Pro Pro Val Asp Lys Glu Ala Glu Ser Tyr 
        195                 200                 205             


Gln Gly Glu Glu Glu Leu Val Ser Gln Thr Asp Val Lys Thr Phe Leu 
    210                 215                 220                 


Glu Ala Leu Pro Gln Glu Ser Trp Gln Asn Lys His Ala Gly Ser Asn 
225                 230                 235                 240 


Gly Asn Arg Ala Thr Asn Gly Ala Pro Ser Leu Val Ser Arg Gly Met 
                245                 250                 255     


Gln Gly Ala Ser Ala Asn Thr Leu Asn Ser Glu Thr Ser Thr Lys Arg 
            260                 265                 270         


Ala Ala Val Tyr Arg Met Asn Arg Asn His Arg Gly Leu Cys Val Ile 
        275                 280                 285             


Val Asn Asn His Ser Phe Thr Ser Leu Lys Asp Arg Gln Gly Thr His 
    290                 295                 300                 


Lys Asp Ala Glu Ile Leu Ser His Val Phe Gln Trp Leu Gly Phe Thr 
305                 310                 315                 320 


Val His Ile His Asn Asn Val Thr Lys Val Glu Met Glu Met Val Leu 
                325                 330                 335     


Gln Lys Gln Lys Cys Asn Pro Ala His Ala Asp Gly Asp Cys Phe Val 
            340                 345                 350         


Phe Cys Ile Leu Thr His Gly Arg Phe Gly Ala Val Tyr Ser Ser Asp 
        355                 360                 365             


Glu Ala Leu Ile Pro Ile Arg Glu Ile Met Ser His Phe Thr Ala Leu 
    370                 375                 380                 


Gln Cys Pro Arg Leu Ala Glu Lys Pro Lys Leu Phe Phe Ile Gln Ala 
385                 390                 395                 400 


Cys Gln Gly Glu Glu Ile Gln Pro Ser Val Ser Ile Glu Ala Asp Ala 
                405                 410                 415     


Leu Asn Pro Glu Gln Ala Pro Thr Ser Leu Gln Asp Ser Ile Pro Ala 
            420                 425                 430         


Glu Ala Asp Phe Leu Leu Gly Leu Ala Thr Val Pro Gly Tyr Val Ser 
        435                 440                 445             


Phe Arg His Val Glu Glu Gly Ser Trp Tyr Ile Gln Ser Leu Cys Asn 
    450                 455                 460                 


His Leu Lys Lys Leu Val Pro Arg Met Leu Lys Phe Leu Glu Lys Thr 
465                 470                 475                 480 


Met Glu Ile Arg Gly Arg Lys Arg Thr Val Trp Gly Ala Lys Gln Ile 
                485                 490                 495     


Ser Ala Thr Ser Leu Pro Thr Ala Ile Ser Ala Gln Thr Pro Arg Pro 
            500                 505                 510         


Pro Met Arg Arg Trp Ser Ser Val Ser 
        515                 520     


<210>  10
<211>  452
<212>  PRT
<213>  Homo sapiens

<400>  10

Met Ala Ala Pro Ser Ala Gly Ser Trp Ser Thr Phe Gln His Lys Glu 
1               5                   10                  15      


Leu Met Ala Ala Asp Arg Gly Arg Arg Ile Leu Gly Val Cys Gly Met 
            20                  25                  30          


His Pro His His Gln Glu Thr Leu Lys Lys Asn Arg Val Val Leu Ala 
        35                  40                  45              


Lys Gln Leu Leu Leu Ser Glu Leu Leu Glu His Leu Leu Glu Lys Asp 
    50                  55                  60                  


Ile Ile Thr Leu Glu Met Arg Glu Leu Ile Gln Ala Lys Val Gly Ser 
65                  70                  75                  80  


Phe Ser Gln Asn Val Glu Leu Leu Asn Leu Leu Pro Lys Arg Gly Pro 
                85                  90                  95      


Gln Ala Phe Asp Ala Phe Cys Glu Ala Leu Arg Glu Thr Lys Gln Gly 
            100                 105                 110         


His Leu Glu Asp Met Leu Leu Thr Thr Leu Ser Gly Leu Gln His Val 
        115                 120                 125             


Leu Pro Pro Leu Ser Cys Asp Tyr Asp Leu Ser Leu Pro Phe Pro Val 
    130                 135                 140                 


Cys Glu Ser Cys Pro Leu Tyr Lys Lys Leu Arg Leu Ser Thr Asp Thr 
145                 150                 155                 160 


Val Glu His Ser Leu Asp Asn Lys Asp Gly Pro Val Cys Leu Gln Val 
                165                 170                 175     


Lys Pro Cys Thr Pro Glu Phe Tyr Gln Thr His Phe Gln Leu Ala Tyr 
            180                 185                 190         


Arg Leu Gln Ser Arg Pro Arg Gly Leu Ala Leu Val Leu Ser Asn Val 
        195                 200                 205             


His Phe Thr Gly Glu Lys Glu Leu Glu Phe Arg Ser Gly Gly Asp Val 
    210                 215                 220                 


Asp His Ser Thr Leu Val Thr Leu Phe Lys Leu Leu Gly Tyr Asp Val 
225                 230                 235                 240 


His Val Leu Cys Asp Gln Thr Ala Gln Glu Met Gln Glu Lys Leu Gln 
                245                 250                 255     


Asn Phe Ala Gln Leu Pro Ala His Arg Val Thr Asp Ser Cys Ile Val 
            260                 265                 270         


Ala Leu Leu Ser His Gly Val Glu Gly Ala Ile Tyr Gly Val Asp Gly 
        275                 280                 285             


Lys Leu Leu Gln Leu Gln Glu Val Phe Gln Leu Phe Asp Asn Ala Asn 
    290                 295                 300                 


Cys Pro Ser Leu Gln Asn Lys Pro Lys Met Phe Phe Ile Gln Ala Cys 
305                 310                 315                 320 


Arg Gly Asp Glu Thr Asp Arg Gly Val Asp Gln Gln Asp Gly Lys Asn 
                325                 330                 335     


His Ala Gly Ser Pro Gly Cys Glu Glu Ser Asp Ala Gly Lys Glu Lys 
            340                 345                 350         


Leu Pro Lys Met Arg Leu Pro Thr Arg Ser Asp Met Ile Cys Gly Tyr 
        355                 360                 365             


Ala Cys Leu Lys Gly Thr Ala Ala Met Arg Asn Thr Lys Arg Gly Ser 
    370                 375                 380                 


Trp Tyr Ile Glu Ala Leu Ala Gln Val Phe Ser Glu Arg Ala Cys Asp 
385                 390                 395                 400 


Met His Val Ala Asp Met Leu Val Lys Val Asn Ala Leu Ile Lys Asp 
                405                 410                 415     


Arg Glu Gly Tyr Ala Pro Gly Thr Glu Phe His Arg Cys Lys Glu Met 
            420                 425                 430         


Ser Glu Tyr Cys Ser Thr Leu Cys Arg His Leu Tyr Leu Phe Pro Gly 
        435                 440                 445             


His Pro Pro Thr 
    450         


<210>  11
<211>  303
<212>  PRT
<213>  Homo sapiens

<400>  11

Met Ala Asp Asp Gln Gly Cys Ile Glu Glu Gln Gly Val Glu Asp Ser 
1               5                   10                  15      


Ala Asn Glu Asp Ser Val Asp Ala Lys Pro Asp Arg Ser Ser Phe Val 
            20                  25                  30          


Pro Ser Leu Phe Ser Lys Lys Lys Lys Asn Val Thr Met Arg Ser Ile 
        35                  40                  45              


Lys Thr Thr Arg Asp Arg Val Pro Thr Tyr Gln Tyr Asn Met Asn Phe 
    50                  55                  60                  


Glu Lys Leu Gly Lys Cys Ile Ile Ile Asn Asn Lys Asn Phe Asp Lys 
65                  70                  75                  80  


Val Thr Gly Met Gly Val Arg Asn Gly Thr Asp Lys Asp Ala Glu Ala 
                85                  90                  95      


Leu Phe Lys Cys Phe Arg Ser Leu Gly Phe Asp Val Ile Val Tyr Asn 
            100                 105                 110         


Asp Cys Ser Cys Ala Lys Met Gln Asp Leu Leu Lys Lys Ala Ser Glu 
        115                 120                 125             


Glu Asp His Thr Asn Ala Ala Cys Phe Ala Cys Ile Leu Leu Ser His 
    130                 135                 140                 


Gly Glu Glu Asn Val Ile Tyr Gly Lys Asp Gly Val Thr Pro Ile Lys 
145                 150                 155                 160 


Asp Leu Thr Ala His Phe Arg Gly Asp Arg Cys Lys Thr Leu Leu Glu 
                165                 170                 175     


Lys Pro Lys Leu Phe Phe Ile Gln Ala Cys Arg Gly Thr Glu Leu Asp 
            180                 185                 190         


Asp Gly Ile Gln Ala Asp Ser Gly Pro Ile Asn Asp Thr Asp Ala Asn 
        195                 200                 205             


Pro Arg Tyr Lys Ile Pro Val Glu Ala Asp Phe Leu Phe Ala Tyr Ser 
    210                 215                 220                 


Thr Val Pro Gly Tyr Tyr Ser Trp Arg Ser Pro Gly Arg Gly Ser Trp 
225                 230                 235                 240 


Phe Val Gln Ala Leu Cys Ser Ile Leu Glu Glu His Gly Lys Asp Leu 
                245                 250                 255     


Glu Ile Met Gln Ile Leu Thr Arg Val Asn Asp Arg Val Ala Arg His 
            260                 265                 270         


Phe Glu Ser Gln Ser Asp Asp Pro His Phe His Glu Lys Lys Gln Ile 
        275                 280                 285             


Pro Cys Val Val Ser Met Leu Thr Lys Glu Leu Tyr Phe Ser Gln 
    290                 295                 300             


<210>  12
<211>  293
<212>  PRT
<213>  Homo sapiens

<400>  12

Met Ser Ser Ala Ser Gly Leu Arg Arg Gly His Pro Ala Gly Gly Glu 
1               5                   10                  15      


Glu Asn Met Thr Glu Thr Asp Ala Phe Tyr Lys Arg Glu Met Phe Asp 
            20                  25                  30          


Pro Ala Glu Lys Tyr Lys Met Asp His Arg Arg Arg Gly Ile Ala Leu 
        35                  40                  45              


Ile Phe Asn His Glu Arg Phe Phe Trp His Leu Thr Leu Pro Glu Arg 
    50                  55                  60                  


Arg Gly Thr Cys Ala Asp Arg Asp Asn Leu Thr Arg Arg Phe Ser Asp 
65                  70                  75                  80  


Leu Gly Phe Glu Val Lys Cys Phe Asn Asp Leu Lys Ala Glu Glu Leu 
                85                  90                  95      


Leu Leu Lys Ile His Glu Val Ser Thr Val Ser His Ala Asp Ala Asp 
            100                 105                 110         


Cys Phe Val Cys Val Phe Leu Ser His Gly Glu Gly Asn His Ile Tyr 
        115                 120                 125             


Ala Tyr Asp Ala Lys Ile Glu Ile Gln Thr Leu Thr Gly Leu Phe Lys 
    130                 135                 140                 


Gly Asp Lys Cys His Ser Leu Val Gly Lys Pro Lys Ile Phe Ile Ile 
145                 150                 155                 160 


Gln Ala Cys Arg Gly Asn Gln His Asp Val Pro Val Ile Pro Leu Asp 
                165                 170                 175     


Val Val Asp Asn Gln Thr Glu Lys Leu Asp Thr Asn Ile Thr Glu Val 
            180                 185                 190         


Asp Ala Ala Ser Val Tyr Thr Leu Pro Ala Gly Ala Asp Phe Leu Met 
        195                 200                 205             


Cys Tyr Ser Val Ala Glu Gly Tyr Tyr Ser His Arg Glu Thr Val Asn 
    210                 215                 220                 


Gly Ser Trp Tyr Ile Gln Asp Leu Cys Glu Met Leu Gly Lys Tyr Gly 
225                 230                 235                 240 


Ser Ser Leu Glu Phe Thr Glu Leu Leu Thr Leu Val Asn Arg Lys Val 
                245                 250                 255     


Ser Gln Arg Arg Val Asp Phe Cys Lys Asp Pro Ser Ala Ile Gly Lys 
            260                 265                 270         


Lys Gln Val Pro Cys Phe Ala Ser Met Leu Thr Lys Lys Leu His Phe 
        275                 280                 285             


Phe Pro Lys Ser Asn 
    290             


<210>  13
<211>  36
<212>  DNA
<213>  artificial

<220>
<223>  1369_Casp3.1_BsmBI_fp

<400>  13
aacgtctctg atgaacagct acaagatgga ctaccc                               36


<210>  14
<211>  37
<212>  DNA
<213>  artificial

<220>
<223>  1369_Casp3.1_2Stop_AscI_rp

<400>  14
aaggcgcgcc tattattttt cgaactgcgg gtggctc                              37


<210>  15
<211>  60
<212>  DNA
<213>  artificial

<220>
<223>  1369_Casp3.1_pHILD2_fp

<400>  15
aacaattgtc tgccatcatg catcatcatc atcatcataa cagctacaag atggactacc     60


<210>  16
<211>  38
<212>  DNA
<213>  artificial

<220>
<223>  1369_Casp3.1_pHILD2_rp

<400>  16
aacaattgtt attagagacc gtggtagaag tacagctc                             38


