1996 Fiscal Year Final Research Report Summary
APPLICATION OF A NOVEL TRYPTASE FOUND IN THE HUMAN RESPIRATORY TRACT FLUID TO THERAPY OF INTERNAL DISEASES
| Project/Area Number |
06557142
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 試験 |
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| Research Field |
応用薬理学・医療系薬学
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| Research Institution | THE UNIVERSITY OF TOKUSHIMA |
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Principal Investigator |
YASUOKA Susumu THE UNIVERSITY OF TOKUSHIMA SCHOOL OF MEDICAL SCIENCES,PROFESSOR, 医療技術短期大学部, 教授 (30035414)
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| Co-Investigator(Kenkyū-buntansha) |
YAMAOKA Kazuyoshi TEIZIN INSTITUTE FOR BIOMEDICAL RESEARCH,THE UNIVERSITY OF TOKUSHIMA RESEARCH FE, 第二創薬研究所, 研究員
MASUDA Ken-ichi TEIZIN INSTITUTE FOR BIOMEDICAL RESEARCH,THE UNIVERSITY OF TOKUSHIMA, 第二創薬研究所, 主任研究員
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| Project Period (FY) |
1994 – 1996
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| Keywords | TRYPSIN-LIKE PROTEASE / SPUTUM / CHRONIC AIRWAY DISEASES / CLONING OF cDNA FOR PROTEASE / RECOMBINANT ENZYME / 組換え酵素 |
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| Research Abstract |
1.PURIFICATION AND CHARACTERIZATION : A NOVEL TRYPSIN-LIKE PROTEASE WITH MOLECULAR WEIGHT OF 27 kDa, WAS ISOLATED FROM SPUTUM OF PATIENTS WITH CHRONIC AIRWAY DISEASES,AND DESIGNATEDAS HUMAN AIRWAY TRYPSIN-LIKE PROTEASE (HAT). THE NH_2-TERMINAL 20 AMINO ACID SEQUENCE OF HAT DID NOT ACCORD WITH THOSE OF ANY KNOWN PROTEASES.
2.CLONING AND CHARACTERIZATION OF cDNA FOR HAT : WE SUCCEEDED IN CLONING OF cDNA FOR HAT FROM TRACHEA DNA.THE PRIMARY STRUCTURE OF HAT WAS DEDUCED FROM THE NUCLEOTIDE SEQUENCE OF THE cDNA.IT WAS SHOWN THAT THE cDNA ENCODE HAT PRECURSOR (MW,47 kDA) WITCH CONSISTED OF NON-CATALYTIC REGION (MW,20kDa) AND CATALYTIC REGION (MW,27kDa), AND THE PRECURSOR WAS TYPE II INTEGRAL MEMBRANE PROTEIN.THE CATALYTIC REGION CORRESPONDED TO HAT,AND WAS THOUGHT TO BE PRODUCED BY LIMITED PROTEOLYSIS OF THE PRECURSOR DURING SECRETION.
3.LOCALIZATION OF HAT : IMMUNOHISTOCHEMICAL STUDY AND NORTHANBLOT ANALYSIS SHOWED THAT HAT IS LOCALIZED MAINLY IN THE SUBMUCOSAL SEROUS GLAND CEELLS.
4.BIOSYNTHESIS OF RECOMBINANT HAT : A RECOMBINANT HAT WAS PRODUCED IN THE INSECT CELLS WITH BACULOVIRUS AS AN VECTOR.
5.BIOLOGICAL ACTIONS OF HAT : BOVINE TRYPSIN ENHANCED INFECTIVITY OF INFLUENZA VIRUS BY CLEAVAGING ITS SURFACE HEMAGLUTININ,BUT HAT DID NOT.FIBRINOGEN,ESPECIALLY ITS alpha-CHAIN WAS CLEAVAGED BY HAT.
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