Chondroitin synthase

Abstract

A vector of the present invention has DNA encoding a protein or a product having the same effect as the protein, the protein containing an amino acid sequence from amino acid numbers 47 to 802 in SEQ. ID. NO:2. Expression of the DNA gives human chondroitin synthase. By using human chondroitin synthase, it is possible to produce a saccharide chain having a repeating disaccharide unit of chondroitin. The DNA or part thereof may be used as a probe for hybridization for the human chondroitin synthase.

1 - 15 . (canceled)  
     
     
         16 . A vector carrying one of DNA (a), (b) or (c), the DNA (b) or (c) encoding a protein having catalytic activities (a) and (i), excluding a DNA encoding a protein at amino-acid position #1 to 802 in SEQ. ID. NO: 2): 
 (a) DNA encoding a protein having an amino acid sequence from amino acid numbers 47 to 802 in SEQ. ID. NO: 2;    (b) DNA that, in a stringent condition, hybridizes with the DNA (a), the DNA complementary with DNA (a),.or DNA having part of a nucleotide sequence of the DNA (a) or the DNA complementary with the DNA (a);    (c) DNA encoding a protein having an amino acid sequence from amino acid numbers 47 to 802 in SEQ. ID. NO:2, wherein one or several amino acids in the amino acid sequence are substituted, deleted, inserted, or transpositioned;    (α) catalytic activity that transferase GalNAc from UDP-GalNAc to chondroitin; 
 (where UDP is uridine 5′diphosphate, and GalNAc is N-acetylgalactosamine residue), and  
   (β) catalytic activity that transferase GlcUA from UDP-GlcUA to chondroitin, 
 (where UDP is uridine 5′diphosphate and GlcUA is N-glucuronic acid residue).  
   
     
     
         17 . The vector as set forth in  claim 16 , wherein: 
 the DNA (a) encodes finding from nucleotide numbers 633 to 2900 in SEQ. ID. NO: 1.    
     
     
         18 . The vector as set forth in  claim 16 , wherein the proteins are soluble.  
     
     
         19 . The vector as set forth in  claim 17 , wherein the proteins are soluble.  
     
     
         20 . The vector as set forth in  claim 16 , being an expression vector.  
     
     
         21 . The vector as set forth in  claim 17 , being an expression vector.  
     
     
         22 . The vector as set forth in  claim 18 , being an expression vector.  
     
     
         23 . The vector as set forth in  claim 19 , being an expression vector.  
     
     
         24 . A transformant whose host is transformed by a vector having any one of DNA (a) to (c), the DNA (b) or (c) encoding a protein having catalytic activities (a) and (f): 
 (a) DNA encoding a protein having an amino acid sequence from amino acid numbers 47 to 802 in SEQ. ID. NO: 2;    (b) DNA that, in a stringent condition, hybridizes with the DNA (a), the DNA complementary with DNA (a), or DNA having part of a nucleotide sequence of the DNA (a) or the DNA complementary with the DNA (a);    (c) DNA encoding a protein having an amino acid sequence from amino acid numbers 47 to 802 in SEQ. ID. NO:2, wherein one or several amino acids in the amino acid sequence are substituted, deleted, inserted, or transpositioned;    (α) catalytic activity that transferase GalNAc from UDP-GalNAc to chondroitin; 
 (where UDP is uridine 5′diphosphate, and GalNAc is N-acetylgalactosamine residue),  
   (β) catalytic activity that transferase from UDP-GlcUA to chondroitin, 
 (where UDP is uridine 5′diphosphate and GlcUA is N-glucuronic acid residue).  
   
     
     
         25 . The transformant as set forth in  claim 24 , wherein: 
 the DNA (a) encodes finding from nucleotide numbers 633 to 2900 in SEQ. ID. NO: 1.    
     
     
         26 . The transformant as set forth in  claim 24 , wherein: 
 the proteins are soluble.    
     
     
         27 . The transformant as set forth in  claim 25 , wherein: 
 the proteins are soluble.    
     
     
         28 . A method for producing chondroitin synthase, the method comprising the steps of: 
 growing a transformant set forth in  claim 24;  and    obtaining the chondroitin synthase from the transformant thus grown.    
     
     
         29 . A method for producing chondroitin synthase, the method comprising the steps of: 
 growing a transformant set forth in  claim 25;  and obtaining the chondroitin synthase from the transformant thus grown.    
     
     
         30 . A method for producing chondroitin synthase, the method comprising the steps of: 
 growing a transformant set forth  claim 26;  and    obtaining the chondroitin synthase from the transformant thus grown.    
     
     
         31 . A method for producing chondroitin synthase, the method comprising the steps of: 
 growing a transformant set forth  claim 27;  and    obtaining the chondroitin synthase from the transformant thus grown.    
     
     
         32 . A reagent for use in chondroitin synthesis, the reagent having an enzyme protein that has an amino acid sequence including an amino acid sequence (A) or (B) and has catalytic activities (α) and (β) 
 (A) amino acid sequence from amino acid numbers 47 to 802 in SEQ. ID. NO: 2;    (B) amino acid sequence from amino acid numbers 47 to 802 in SEQ. ID. NO:2, wherein one or several amino acids in the amino acid sequence are substituted, deleted, inserted, or transpositioned.    (α) catalytic activity that transferase GalNAc from UDP-GalNAc to chondroitin, 
 (where UDP is uridine 5′diphosphate, and GalNAc is N-acetylgalactosamine residue);  
   (β) catalytic activity that transferase GlcUA from UDP-GlcUA to chondroitin, 
 (where UDP is uridine 5′diphosphate and GlcUA is glucuronic acid residue).  
   
     
     
         33 . The reagent as set forth in  claim 32 , wherein the enzyme protein is soluble.  
     
     
         34 . A method for producing a saccharide chain expressed by Formula (3), the method comprising at least the step of causing a reagent to contact with GalNAc donor and a saccharide chain expressed by Formula (1), the reagent set forth in  claim 32:   
         GlcUA-GalNAc-R 1    (1),  GalNAc-GlcUA-GalNAc-R 1    (3),  
       (where GlcUA and GalNAc are as defined above, “-” indicates a glycoside linkage, R 1  is an arbitrary group).  
     
     
         35 . A method for producing a saccharide chain expressed by Formula (3), the method comprising at least the step of causing a reagent to contact with GalNAc donor and a saccharide chain expressed by Formula (1), the reagent set forth in  claim 33:   
         GlcUA-GalNAc-R 1    (1),  GalNAc-GlcUA-GalNAc-R 1    (3),  
       (where GlcUA and GalNAc are as defined above, “-” indicates a glycoside linkage, R 1  is an arbitrary group)  
     
     
         36 . A method for producing a saccharide chain expressed by Formula (4), the method comprising at least the step of causing a reagent to contact with GlcUA donor and a saccharide chain expressed by Formula (2), the reagent set forth in  claim 32:   
         GalNAc-GlcUA-R 2    (2),  GlcUA-GalNAc-GlcUA-R 2    (4),  
       (where GlcUA, GalNAc, and “-” are as defined above, R 2  is an arbitrary group).  
     
     
         37 . A method for producing a saccharide chain expressed by Formula (4), the method comprising at least the step of causing a reagent to contact with GlcUA donor and a saccharide chain expressed by Formula (2), the reagent set forth in  claim 33:   
         GalNAc-GlcUA-R 2 ( 2),  GlcUA-GalNAc-GlcUA-R 2    (4),  
       (where GlcUA, GalNAc, and “-” are as defined above, R 2  is an arbitrary group).  
     
     
         38 . A method for producing a saccharide chain selected from saccharide chains expressed by Formulas (5) and (7) respectively, the method comprising at least the step of causing a reagent to contact with GalNAc donor, GlcUA donor and a saccharide chain expressed by Formula (1), the reagent set forth in  claim 32:   
         GlcUA-GalNAc-R 1    (1),  (GlcUA-GalNAc)n-GlcUA-GalNAc-R 1    (5),  GalNAc-(GlcUA-GalNAc)n-GlcUA-GalNAc-R 1    (7),  
       (where n is an integer not less than 1, GlcUA, GalNAc, and “-” are as defined above, R 1  is an arbitrary group).  
     
     
         39 . A method for producing a saccharide chain selected from saccharide chains expressed by Formulas (5) and (7) respectively, the method comprising at least the step of causing a reagent to contact with GalNAc donor, GlcUA donor and a saccharide chain expressed by Formula (1), the reagent set forth in  claim 33:   
         GlcUA-GalNAc-R 1    (1),  (GlcUA-GalNAc)n-GlcUA-GalNAc-R 1    (5),  GalNAc-(GlcUA-GalNAc)n-GlcUA-GalNAc-R 1    (7),  
       (where n is an integer not less than 1, GlcUA, GalNAc, and “-” are as defined above, R 1  is an arbitrary group).  
     
     
         40 . A method for producing a saccharide chain selected from saccharide chains expressed by Formulas (6) and (8) respectively, the method comprising at least the step of causing a reagent to contact with GalNAc donor, GlcUA donor and a saccharide chain expressed by Formula (2), the reagent set forth in  claim 32:   
         GalNAc-GlcUA-R 2    (2),  (GalNAc-GlcUA)n-GalNAc-GlcUA-R 2    (6),  GlcUA-(GalNAc-GlcUA)n-GalNAc-GlcUA-R 2    (8),  
       (where n is an integer not less than 1, GlcUA, GalNAc, and “-” are as defined above, R 2  is an arbitrary group).  
     
     
         41 . A method for producing a saccharide chain selected from saccharide chains expressed by Formulas (6) and (8) respectively, the method comprising at least the step of causing a reagent to contact with GalNAc donor, GlcUA donor and a saccharide chain expressed by Formula (2), the reagent set forth in  claim 33:   
         GalNAc-GlcUA-R 2 ( 2),  (GalNAc-GlcUA)n-GalNAc-GlcUA-R 2    (6),  GlcUA-(GalNAc-GlcUA)n-GalNAc-GlcUA-R 2    (8),  
       (where n is an integer not less than 1, GlcUA, GalNAc, and “-” are as defined above, R 2  is an arbitrary group).  
     
     
         42 . A probe for hybridization, the probe containing a nucleotide sequence from nucleotide numbers 495 to 2900 in SEQ. ID. NO: 1, or a sequence complementary with part of the nucleotide sequence.