1988 Fiscal Year Final Research Report Summary
Analytical Studies of Plasma Membrane Biogenesis in Transporting Epithelia
| Project/Area Number |
61480127
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Pathological medical chemistry
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| Research Institution | Hamamatsu University School of Medicine |
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Principal Investigator |
FUJITA Michiya Hamamatsu University School of Medicine Professor, 医学部, 教授 (60014031)
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| Co-Investigator(Kenkyū-buntansha) |
TAKESUE Yoshiki Gifu University Faculty Faculty of Liberal Arts Associate Professor, 教養部, 助教授 (50023651)
UEZATO Tadayoshi Hamamatsu University School of Medicine Professor, 医学部, 助手 (40115465)
OHTA Hidehiko Hamamatsu University School of Medicine Professor, 医学部, 助教授 (00014192)
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| Project Period (FY) |
1986 – 1988
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| Keywords | Enterocyte / Renal tubular epithelial cell / Glucose transporter / Na,K-ATPase / Trehalase / Cytochalasin B / Membrane protein / 蛍光抗体 |
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| Research Abstract |
1.Microvullus membrane(MVM) and basolateral membrane (BLM) were prepared from small intestinal epithelial cells and were characterized biochemically and morphologically.
2.Glucose transport by MVM vesicles was inhibited cytochalasin B (CYB). 52K protein was photolabeled by CYB. Glucose transport by MVM vesicles was inhibited maximally 60% by CYB (Ki=10 M) and 86K protein was labeled with CYB (Kd=8 M,Bmax=70pmol/mg protein).
3.Fluorescent antibody against 70K MVM protein reacted not only with intestinal MVM but with renal and hepatic MVM. Antibody against Na,K-ATPasereacted with intestinal, renal and hepatic BLM.
4.Trehalase of renal MVM turned out to be covalently bound with MVM viaphosphatidyl inositol. Trehalasesolubilized with Triton X-100 was hydrophobic and adsorbed to phenyl sepharose clumn. Iucubated at 37 c, it became hydrophilic and this conversion was not depressed by any protease inhibitors so far known.
6.Bioassembly of MVM and BLM proteins showed monophasic time course for MVM and biphasic one for BLM. Possible contamination of BLM with endoplasmic reticulum membrane is under investigation to account for discrepancy.
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