| Project/Area Number |
12680610
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Structural biochemistry
|
|---|
| Research Institution | Aichi University of Education |
|---|
Principal Investigator |
HABUCHI Osami Aichi University of Education, Science, Professor, 教育学部, 教授 (90024067)
|
|---|
| Project Period (FY) |
2000 – 2001
|
| Project Status |
Completed (Fiscal Year 2001)
|
| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2001: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2000: ¥2,100,000 (Direct Cost: ¥2,100,000)
|
|---|
| Keywords | Chondroitin 4-sulfotransferase / GalNAc 4-sulfate 6-O-sulfotransferase / Chondroitin sulfate E / Squid cartilage / GalNAc 4-sulfotransferase / Chondroitin 6-sulfotransferase / GalNAc4硫酸6硫酸転移酵素 / GalNAc4硫酸転移酵素 / シピレエイ |
|---|
| Research Abstract |
(1) We cloned mouse and human chondroitin 4-sulfotransferase gene, characterized the protein expressed from the gene and determined expression pattern in various tissues. We also mapped the gene in human chromosome.
(2) We cloned and characterized human GalNAc 4-sulfotransferase that strongly expressed in human pituitary grand. We also cloned mouse GalNAc 4-sulfotransferase I and II and determined in various tissues by Northern blot and RT-PCR.
(3) We expressed NSIST, which was cloned from Torpedo electric organ, in COS-7 cells and found that substrate specificity of NSIST was very similar to that of chondroitin 6-sulfotransferase.
(4) Af-Acetylgalactosamine 4-sulfate 6-O-sulfotransferase (GalNAc4S-6ST)was purified to apparent homogeneity from the squid cartilage. The purified enzyme transferred sulfate from PAPS to chondroitin sulfate A, chondroitin sulfate C and dermatan sulfate. The transfer of sulfate to chondroitin sulfate A and dermatan sulfate occurred mainly at position 6 of the i
… More
nternal Af-acetylgalactosamine 4-sulfate residues. When a trisaccharide or a pentasaccharide having sulfate groups at position 4 of W-acetylgalactosamine residues were used as acceptors, efficient sulfation of position 6 at nonreducing terminal N-acetylgalactosamine 4-sulfate residue was observed. We cloned and characterized human GalNAc4S-6ST. The strategy for identification of human GalNAc4S-6ST consisted of: 1) determination of the amino acid sequences of peptides derived from the purified squid GalNAc4S-6ST, 2) amplification of squid DNA by PCR, and 3) homology search using the amino acid sequence deduced from the squid DNA. The recombinant protein expressed from the human GalNAc4S-6ST cDNA transferred sulfate from PAPS to position 6 of the nonreducing terminal and internal GalNAc (4SO_4) residues contained in chondroitin sulfate A and dermatan sulfate. When a trisaccharide and a pentasaccharide having sulfate groups at position 4 of N-acetylgalactosamine residues were used as acceptors, only nonreducing terminal GalNAc (4SO_4) residues were sulfated. The nucleotide sequence of the human GalNAc4S-6ST cDNA was nearly identical to the sequence of human B cell RAG-associated gene. Less
|
