1997 Fiscal Year Final Research Report Summary
Biochemical Study on Novel Bioregulatory Peptides
| Project/Area Number |
07406005
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Bioproduction chemistry/Bioorganic chemistry
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| Research Institution | University of Tsukuba |
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Principal Investigator |
GOTO Katsutoshi University of Tsukuba, Institute of Basic Medical Scienses.Professor, 応用生物化学系, 教授 (60072766)
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| Co-Investigator(Kenkyū-buntansha) |
MUKAI Hidehito University of Tsukuba, Institute of Applied Biochemistry.Assistant Professor, 応用生物化学系, 講師 (20251027)
後藤 勝年 筑波大学, 基礎医学系, 教授 (30012660)
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| Project Period (FY) |
1995 – 1997
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| Keywords | Regulatory Peptide / Cytokine / Chemokine / Neurtrophil Activator / Survival Factor / Structure-Analysis / Amino Acid Sequence / Mass-Spectra |
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| Research Abstract |
It is considered that much more unknown endogenous bioregulatory peptide or small protein exist in mammalian organs. To find out novel bioregulatory peptide or protein and disclose physiological role of them are expected to provide not only significant knowledge to understand precise physiological function basically but also give useful information for sustaining health and development for guideline of creating new drug or therapeutic method.
The aim of this research project was the systematic screening of the peptide or protein from porcine heart which involved in bioregulation or bioprotection of the circulation system around the heart in mammals. The isolation of novel peptide or protein factor, will be followed by the structural determination and synthesis by chemical method or with the aid of molecular biology technique and study the biological function from physiological and pharmacological viewpoints.
In this research, thetarget of screening was focussed to novel peptide factor related to so called "chemokine" and bioassey systems used were (1) beta-hexosaminidase secretion by the activation of neutrophil-like differetiated HL-60 (Human eukemia 60) cell and (2) survival activity in a fibroblastoma NIH/3T3 cell using MTT method.
From the diluted acetic acid extracts of porcine heart, relatively low molecular protein fractions which showed activity in the assay described above were collected and purified and the structure of the peptidergic factor were analyzed by amino acid sequencing and high resolution mass spectrometry. After the structure determination, the peptides were synthesized chemically and molecular biologically.
As a first result, active peptide which showed strong activating potency to neutrophil-like cell was identified as amino terminal portion of porcine cytochrome b. The second, a novel peptide with 62 amino acids bearing three disulfide bridges was isolated as survival principle in NIH/3T3 cell system.
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