2001 Fiscal Year Final Research Report Summary
DEVELOPMENT OF VEROTOXIN INHIBITOR BY USING HYBRID-LIBRARY THCHNIQUES
| Project/Area Number |
11672236
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Environmental pharmacy
|
|---|
| Research Institution | Research Institute, International Medical Center of Japan |
|---|
Principal Investigator |
NOSHIKAWA Kiyotaka INTERNATIONA MEDICAL CENTER OF JAPAN, RESEARCH INSTITUTE, DEPARTMENT OF CLINICAL PHARMACOLOGY, DIVISION HEAD, 研究所・臨床薬理研究部, 室長 (40218128)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
NATORI Yasuhiro INTERNATIONA MEDICAL CENTER OF JAPAN, RESEARCH INSTITUTE, DEPARTMENT OF CLINICAL PHARMACOLOGY, DIRECTOR, 研究所・臨床薬理研究部, 部長 (10164485)
|
|---|
| Project Period (FY) |
1999 – 2000
|
| Keywords | PEPTIDE LIBRARY / VEROTOXIN / INHIBITOR / AMINO ACID ANALOG |
|---|
| Research Abstract |
1) Preparation of hybrid peptide library ; Two amino acid analogs, in which Galα1-OH or Galβ1-OH present in Gb3 (Galα1-4 Galβ1-4 Glvα1-ceramide, receptor for verotoxin) bind to the OH-group of Ser, were prepared by Oglicoside-condensation reaction. Hybrid-peptide library was synthesized by using one of the above synthetic amino acid analogs.
2) Determination of a high affinity binding motif for verotoxin by using hybrid-peptide library ; At first, in order to see whether a high affinity binding motif for immobilized enzyme could be determined by using peptide library technique, ZAP-70, a Tyr kinase, was used. We found that a high affinity and highly selective binding motif for ZAP-70 could be determined by this technique (ref. 1). We prepared glutathione-S-transferase (GST) fused verotoxin affinity column, and performed screening of high affinity binding motif for verotoxin by using hybrid-peptide library. We found that acidic amino acid Glu was selected at positions +3 and +4, and Val was selected at position +1 (position 0 corresponds to the position of amino acid analog present in the hybrid-peptide library). It is speculated that this motif could bind to the site I present in B-subunits of verotoxin by judging from the reported crystal structure of B-subunits with sugar moiety of Gb3.Determination of a binding motif of the amino terminus side is undergoing.
|
