| Project/Area Number |
07670670
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Respiratory organ internal medicine
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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) |
SANO Toshiaki THE UNIVERSITY OF TOKUSHIMA SCHOOL OF MEDICINE,PROFESSOR, 医学部, 教授 (80154128)
KOYAMA Hajime THE UNIVERSITY OF TOKUSHIMA SCHOOL OF MEDICINE,ASSISTANT PROFESSOR, 医学部, 助教授 (80109074)
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1996: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1995: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | TRYPSIN-LIKE PROTEASE / SPUTUM / CHRONIC AIRWAY DISEASES / CLONING OF cDNA FOR PROTEASE / RECOMBINANT ENZYME / INFULENZA VIRUS / インフルエンザウイルス / ヘマアグルチニン / 気道トリプターゼ / 喀痰 / 気道トリプターゼ遺伝子 / 気道粘膜下漿液腺 / リコンビナントトリプターゼ |
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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 DESIGNATED AS 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 NORTHAN BLOT 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 : IN VITRO,BOVINE TRYPSIN ENHANCED INFECTIVITY OF INFLUENZA VIRUS BY CLEAVAGING ITS SURFACE HEMAGLUTININ,BUT HAT DID NOT.THE SUBSTRATE SPECIFICY OF HAT WAS CHANGED BY VARIOUS CONDITIONS,SUCH AS EXISTENCE OF SALTS AND ITS ORGANIC SOLVENT TREATMENT.THE REFORE FURTHER STUDY IS NECESSARY TO CLARIFY PATHOPHYSIOLOGOCAL SIGNIFICANCE OF HAT IN INFLUENZA VIRUS INFECTION IN THE AIRWAY.
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