| Project/Area Number |
17570094
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Structural biochemistry
|
|---|
| Research Institution | KYUSHU UNIVERSITY |
|---|
Principal Investigator |
KAKUTA Yoshimitsu Kyushu University, Faculty of Agriculture, Associate Professor, 農学研究院, 助教授 (00314360)
|
|---|
| Project Period (FY) |
2005 – 2006
|
| Project Status |
Completed (Fiscal Year 2006)
|
| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2006: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 2005: ¥1,800,000 (Direct Cost: ¥1,800,000)
|
|---|
| Keywords | Sulfotransferase / substrate / crystallography / X線結晶構造解析 / 構造生物学 |
|---|
| Research Abstract |
We have determined crystal structures of sulfotransferase STF1 from Mycobacterium tuberculosis H37Rv at 1.5Å. The predicted amino acid sequence of STF1 from genomic sequence of M. tuberculosis have homology to mammalian cytosolic sulfotransferases only the region for PAPS binding sequence motif. STF1 was crystallized in two conditions: neutral (pH 7.1) and acidic (pH 4.6) with 1M sulfobetaine. Diffractions of these crystals were observed with synchrotron radiation at SPring-8. The phases were determined with MIRAS method for acidic condition crystal and with molecular replacement method for the neutral condition crystal using acidic condition structure. These structures are almost identical but the predicted acceptor substrate binding region was denatured in the structure from acidic condition. The STF1 structure had 5-stranded parallel beta sheet with PAPS binding motifs (5'-PSB loop and 3'-PB motif) which are typically conserved among all known sulfotransferase structures. It has unique structures at the amino terminal which forms 3-helical alpha-bundle. We observed STF1 have specific affinity to maltose with PAP using surface plasmon resonance (SPR) detection. The predicted substrate-binding site of STF1 had small pocket suitable for small molecules such as maltose. These results suggest that STF1 may be associated on biosynthesis of sulfated product including carbohydrate and glycolipid.
|
