
SEQUENCE LISTING

<110> General Atomics
      Yuan, Chong-Sheng
      Datta, Abhijit
      Wang, Yuping
      

<120> METHODS AND COMPOSITIONS FOR
      DETERMINATION OF GLYCATED PROTEINS
  

<130> 466992001340

<140> To Be Assigned
<141> Filed Herewith	

<150> 10/622,893          
<151> 2003-07-17  

<160> 23

<170> FastSEQ for Windows Version 4.0

<210> 1
<211> 12
<212> PRT
<213> Artificial Sequence

<220> 
<223> 40%-100% identity to leader sequence

<400> 1
Met Gly Gly Ser Gly Asp Asp Asp Asp Leu Ala Leu
 1               5                  10          


<210> 2
<211> 6
<212> PRT
<213> Artificial Sequence

<220> 
<223> FAD cofactor-binding consensus sequence
      

<221> VARIANT        
<222> 2, 4, 5
<223> Xaa = Any Amino Acid

<400> 2
Gly Xaa Gly Xaa Xaa Gly
 1               5      


<210> 3
<211> 437
<212> PRT
<213> Artificial Sequence

<220> 
<223> 40%-100% identity to the amadoriase
      

<400> 3
Ala Val Thr Lys Ser Ser Ser Leu Leu Ile Val Gly Ala Gly Thr Trp
 1               5                  10                  15      
Gly Thr Ser Thr Ala Leu His Leu Ala Arg Arg Gly Tyr Thr Asn Val
            20                  25                  30          
Thr Val Leu Asp Pro Tyr Pro Val Pro Ser Ala Ile Ser Ala Gly Asn
        35                  40                  45              
Asp Val Asn Lys Val Ile Ser Ser Gly Gln Tyr Ser Asn Asn Lys Asp
    50                  55                  60                  
Glu Ile Glu Val Asn Glu Ile Leu Ala Glu Glu Ala Phe Asn Gly Trp
65                  70                  75                  80  
Lys Asn Asp Pro Leu Phe Lys Pro Tyr Tyr His Asp Thr Gly Leu Leu
                85                  90                  95      
Met Ser Ala Cys Ser Gln Glu Gly Leu Asp Arg Leu Gly Val Arg Val
            100                 105                 110         
Arg Pro Gly Glu Asp Pro Asn Leu Val Glu Leu Thr Arg Pro Glu Gln
        115                 120                 125             
Phe Arg Lys Leu Ala Pro Glu Gly Val Leu Gln Gly Asp Phe Pro Gly
    130                 135                 140                 
Trp Lys Gly Tyr Phe Ala Arg Ser Gly Ala Gly Trp Ala His Ala Arg
145                 150                 155                 160 
Asn Ala Leu Val Ala Ala Ala Arg Glu Ala Gln Arg Met Gly Val Lys
                165                 170                 175     
Phe Val Thr Gly Thr Pro Gln Gly Arg Val Val Thr Leu Ile Phe Glu
            180                 185                 190         
Asn Asn Asp Val Lys Gly Ala Val Thr Gly Asp Gly Lys Ile Trp Arg
        195                 200                 205             
Ala Glu Arg Thr Phe Leu Cys Ala Gly Ala Ser Ala Gly Gln Phe Leu
    210                 215                 220                 
Asp Phe Lys Asn Gln Leu Arg Pro Thr Ala Trp Thr Leu Val His Ile
225                 230                 235                 240 
Ala Leu Lys Pro Glu Glu Arg Ala Leu Tyr Lys Asn Ile Pro Val Ile
                245                 250                 255     
Phe Asn Ile Glu Arg Gly Phe Phe Phe Glu Pro Asp Glu Glu Arg Gly
            260                 265                 270         
Glu Ile Lys Ile Cys Asp Glu His Pro Gly Tyr Thr Asn Met Val Gln
        275                 280                 285             
Ser Ala Asp Gly Thr Met Met Ser Ile Pro Phe Glu Lys Thr Gln Ile
    290                 295                 300                 
Pro Lys Glu Ala Glu Thr Arg Val Arg Ala Leu Leu Lys Glu Thr Met
305                 310                 315                 320 
Pro Gln Leu Ala Asp Arg Pro Phe Ser Phe Ala Arg Ile Cys Trp Cys
                325                 330                 335     
Ala Asp Thr Ala Asn Arg Glu Phe Leu Ile Asp Arg His Pro Gln Tyr
            340                 345                 350         
His Ser Leu Val Leu Gly Cys Gly Ala Ser Gly Arg Gly Phe Lys Tyr
        355                 360                 365             
Leu Pro Ser Ile Gly Asn Leu Ile Val Asp Ala Met Glu Gly Lys Val
    370                 375                 380                 
Pro Gln Lys Ile His Glu Leu Ile Lys Trp Asn Pro Asp Ile Ala Ala
385                 390                 395                 400 
Asn Arg Asn Trp Arg Asp Thr Leu Gly Arg Phe Gly Gly Pro Asn Arg
                405                 410                 415     
Val Met Asp Phe His Asp Val Lys Glu Trp Thr Asn Val Gln Tyr Arg
            420                 425                 430         
Asp Ile Ser Lys Leu
        435         


<210> 4
<211> 17
<212> PRT
<213> Artificial Sequence

<220> 
<223> 40%-100% identity of the second bacterial leader sequence
      

<400> 4
Lys Gly Glu Leu Glu Gly Leu Pro Ile Pro Asn Pro Leu Leu Arg Thr
 1               5                  10                  15      
Gly
    


<210> 5
<211> 472
<212> PRT
<213> Artificial Sequence

<220> 
<223> chimeric protein
      

<400> 5
Met Gly Gly Ser Gly Asp Asp Asp Asp Leu Ala Leu Ala Val Thr Lys
 1               5                  10                  15      
Ser Ser Ser Leu Leu Ile Val Gly Ala Gly Thr Trp Gly Thr Ser Thr
            20                  25                  30          
Ala Leu His Leu Ala Arg Arg Gly Tyr Thr Asn Val Thr Val Leu Asp
        35                  40                  45              
Pro Tyr Pro Val Pro Ser Ala Ile Ser Ala Gly Asn Asp Val Asn Lys
    50                  55                  60                  
Val Ile Ser Ser Gly Gln Tyr Ser Asn Asn Lys Asp Glu Ile Glu Val
65                  70                  75                  80  
Asn Glu Ile Leu Ala Glu Glu Ala Phe Asn Gly Trp Lys Asn Asp Pro
                85                  90                  95      
Leu Phe Lys Pro Tyr Tyr His Asp Thr Gly Leu Leu Met Ser Ala Cys
            100                 105                 110         
Ser Gln Glu Gly Leu Asp Arg Leu Gly Val Arg Val Arg Pro Gly Glu
        115                 120                 125             
Asp Pro Asn Leu Val Glu Leu Thr Arg Pro Glu Gln Phe Arg Lys Leu
    130                 135                 140                 
Ala Pro Glu Gly Val Leu Gln Gly Asp Phe Pro Gly Trp Lys Gly Tyr
145                 150                 155                 160 
Phe Ala Arg Ser Gly Ala Gly Trp Ala His Ala Arg Asn Ala Leu Val
                165                 170                 175     
Ala Ala Ala Arg Glu Ala Gln Arg Met Gly Val Lys Phe Val Thr Gly
            180                 185                 190         
Thr Pro Gln Gly Arg Val Val Thr Leu Ile Phe Glu Asn Asn Asp Val
        195                 200                 205             
Lys Gly Ala Val Thr Gly Asp Gly Lys Ile Trp Arg Ala Glu Arg Thr
    210                 215                 220                 
Phe Leu Cys Ala Gly Ala Ser Ala Gly Gln Phe Leu Asp Phe Lys Asn
225                 230                 235                 240 
Gln Leu Arg Pro Thr Ala Trp Thr Leu Val His Ile Ala Leu Lys Pro
                245                 250                 255     
Glu Glu Arg Ala Leu Tyr Lys Asn Ile Pro Val Ile Phe Asn Ile Glu
            260                 265                 270         
Arg Gly Phe Phe Phe Glu Pro Asp Glu Glu Arg Gly Glu Ile Lys Ile
        275                 280                 285             
Cys Asp Glu His Pro Gly Tyr Thr Asn Met Val Gln Ser Ala Asp Gly
    290                 295                 300                 
Thr Met Met Ser Ile Pro Phe Glu Lys Thr Gln Ile Pro Lys Glu Ala
305                 310                 315                 320 
Glu Thr Arg Val Arg Ala Leu Leu Lys Glu Thr Met Pro Gln Leu Ala
                325                 330                 335     
Asp Arg Pro Phe Ser Phe Ala Arg Ile Cys Trp Cys Ala Asp Thr Ala
            340                 345                 350         
Asn Arg Glu Phe Leu Ile Asp Arg His Pro Gln Tyr His Ser Leu Val
        355                 360                 365             
Leu Gly Cys Gly Ala Ser Gly Arg Gly Phe Lys Tyr Leu Pro Ser Ile
    370                 375                 380                 
Gly Asn Leu Ile Val Asp Ala Met Glu Gly Lys Val Pro Gln Lys Ile
385                 390                 395                 400 
His Glu Leu Ile Lys Trp Asn Pro Asp Ile Ala Ala Asn Arg Asn Trp
                405                 410                 415     
Arg Asp Thr Leu Gly Arg Phe Gly Gly Pro Asn Arg Val Met Asp Phe
            420                 425                 430         
His Asp Val Lys Glu Trp Thr Asn Val Gln Tyr Arg Asp Ile Ser Lys
        435                 440                 445             
Leu Lys Gly Glu Leu Glu Gly Leu Pro Ile Pro Asn Pro Leu Leu Arg
    450                 455                 460                 
Thr Gly His His His His His His
465                 470         


<210> 6
<211> 1419
<212> DNA
<213> Artificial Sequence

<220> 
<223> nucleotide sequence encoding a chimeric protein
      

<400> 6
atgggaggtt cgggtgacga tgatgacctg gctctcgccg tcactaagtc atcatctctc 60
ctgatcgttg gtgccgggac ttggggcacc tcaacggctc tgcacctcgc gcgccgcgga 120
tataccaacg ttaccgtgct ggacccctat cctgtcccta gcgccatctc cgccggaaac 180
gacgtgaaca aagtcattag cagtggccaa tattcgaata acaaagacga aatcgaagtg 240
aatgagatct tggcggaaga ggcgtttaac ggttggaaga acgacccgct tttcaaaccg 300
tattatcatg atacgggcct gctgatgtct gcttgctcgc aggagggcct ggatcgcctg 360
ggcgtccggg tacgtccggg cgaggatcct aatctggtgg aacttacccg cccggagcaa 420
tttcgtaaac tggccccgga aggcgtgttg caaggtgatt ttccgggttg gaaagggtac 480
tttgcgcgtt ccggcgctgg ctgggcacat gcaaggaatg ccttagtggc agcagcacgc 540
gaagcacagc gcatgggtgt aaaatttgtt actggcaccc cgcagggtcg tgtagtcacg 600
ttaatctttg aaaataacga tgtaaaaggt gccgttacgg gcgatggcaa aatttggaga 660
gcggaacgta cattcctgtg tgctggggct agcgcgggtc agttcctaga tttcaagaat 720
caacttcgac caaccgcttg gaccctggta cacattgcgt taaaaccgga agaacgtgcg 780
ttgtacaaaa atataccggt tatctttaac atcgaacggg ggtttttctt tgaacccgat 840
gaggagcgcg gtgagattaa aatatgcgat gaacacccgg gctacacaaa tatggtccag 900
agtgcagacg gcacgatgat gagcattccg ttcgaaaaaa cccagattcc aaaagaagcc 960
gaaacgcgcg ttcgggccct gctgaaagag acaatgcccc agctggcaga ccgtccattc 1020
agcttcgcac gcatttgctg gtgtgccgat accgcgaatc gcgaattcct gatagatcga 1080
catccgcagt accacagtct tgtgttgggc tgtggtgcga gcggaagagg gtttaaatat 1140
ctgccttcta ttgggaatct cattgttgac gcgatggaag gtaaagtgcc gcaaaaaatt 1200
cacgaattaa tcaagtggaa cccggacatt gcggcgaacc gtaactggcg tgatactctg 1260
gggcgttttg gcggtccaaa tcgtgtgatg gattttcatg atgtgaagga atggaccaat 1320
gttcagtatc gtgatatttc caagctgaaa ggagagttgg aaggtaagcc aatccctaac 1380
ccgttactgc gcacaggcca tcaccatcat catcattaa                        1419

<210> 7
<211> 39
<212> PRT
<213> Artificial Sequence

<220> 
<223> sequence homology between the N-terminal sequence of Amadoriases Ia
      

<221> VARIANT        
<222> 12
<223> Xaa = C or T

<400> 7
Ala Pro Ser Ile Leu Ser Thr Glu Ser Ser Ile Xaa Val Ile Gly Ala
 1               5                  10                  15      
Gly Thr Trp Gly Gly Gly Gly Gly Gly Gly Gly Gly Gly Gly Gly Gly
            20                  25                  30          
Gly Gly Gly Gly Gly Gly Gly
        35                  


<210> 8
<211> 39
<212> PRT
<213> Artificial Sequence

<220> 
<223> Sequence homology between the N-terminal sequence of Amadoriase Ib
      

<400> 8
Ala Pro Ser Ile Leu Ser Thr Glu Ser Ser Ile Ile Val Ile Gly Ala
 1               5                  10                  15      
Gly Thr Trp Gly Gly Gly Gly Gly Gly Gly Gly Gly Gly Gly Gly Gly
            20                  25                  30          
Gly Gly Gly Gly Gly Gly Gly
        35                  


<210> 9
<211> 39
<212> PRT
<213> Artificial Sequence

<220> 
<223> Sequence homology between the N-terminal sequence of Amadoriase Ic
      

<400> 9
Ser Thr Glu Ser Ser Ile Ile Val Ile Gly Ala Gly Thr Trp Gly Cys
 1               5                  10                  15      
Ser Thr Ala Leu Leu Leu Leu Leu Leu Leu Leu Leu Leu Leu Leu Leu
            20                  25                  30          
Leu Leu Leu Leu Leu Leu Leu
        35                  


<210> 10
<211> 39
<212> PRT
<213> Artificial Sequence

<220> 
<223> Sequence homology between the N-terminal sequence of Amadoriase II
      

<400> 10
Ala Val Thr Lys Ser Ser Ser Leu Leu Ile Val Gly Ala Gly Thr Trp
 1               5                  10                  15      
Gly Thr Ser Thr Thr Thr Thr Thr Thr Thr Thr Thr Thr Thr Thr Thr
            20                  25                  30          
Thr Thr Thr Thr Thr Thr Thr
        35                  


<210> 11
<211> 7
<212> PRT
<213> Artificial Sequence

<220> 
<223> exemplary epitope tag
      

<400> 11
Asp Tyr Lys Asp Asp Asp Lys
 1               5          


<210> 12
<211> 9
<212> PRT
<213> Artificial Sequence

<220> 
<223> exemplary epitope tag
      

<400> 12
Tyr Pro Tyr Asp Val Pro Asp Tyr Ala
 1               5                  


<210> 13
<211> 11
<212> PRT
<213> Artificial Sequence

<220> 
<223> exemplary epitope tag
      

<400> 13
Cys Gln Asp Leu Pro Gly Asn Asp Asn Ser Thr
 1               5                  10      


<210> 14
<211> 10
<212> PRT
<213> Artificial Sequence

<220> 
<223> exemplary epitope tag
      

<400> 14
Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu
 1               5                  10  


<210> 15
<211> 6
<212> PRT
<213> Artificial Sequence

<220> 
<223> exemplary epitope tag
      

<400> 15
His His His His His His
 1               5      


<210> 16
<211> 6
<212> PRT
<213> Artificial Sequence

<220> 
<223> exemplary epitope tag
      

<400> 16
Asp Thr Tyr Arg Tyr Ile
 1               5      


<210> 17
<211> 6
<212> PRT
<213> Artificial Sequence

<220> 
<223> exemplary epitope tag
      

<400> 17
Glu Tyr Met Pro Met Glu
 1               5      


<210> 18
<211> 11
<212> PRT
<213> Artificial Sequence

<220> 
<223> exemplary epitope tag
      

<400> 18
Ala Ser Met Thr Gly Gly Gln Gln Met Gly Arg
 1               5                  10      


<210> 19
<211> 10
<212> PRT
<213> Artificial Sequence

<220> 
<223> exemplary epitope tag
      

<400> 19
Ser Phe Pro Gln Phe Lys Pro Gln Glu Ile
 1               5                  10  


<210> 20
<211> 12
<212> PRT
<213> Artificial Sequence

<220> 
<223> exemplary epitope tag
      

<400> 20
Lys Gly Phe Ser Tyr Phe Gly Glu Asp Leu Met Pro
 1               5                  10          


<210> 21
<211> 6
<212> PRT
<213> Artificial Sequence

<220> 
<223> exemplary epitope tag
      

<400> 21
Gln Tyr Pro Ala Leu Thr
 1               5      


<210> 22
<211> 11
<212> PRT
<213> Artificial Sequence

<220> 
<223> exemplary epitope tag
      

<400> 22
Gln Arg Gln Tyr Gly Asp Val Phe Lys Gly Asp
 1               5                  10      


<210> 23
<211> 10
<212> PRT
<213> Artificial Sequence

<220> 
<223> exemplary epitope tag
      

<400> 23
Glu Val His Thr Asn Gln Asp Pro Leu Asp
 1               5                  10  




