1995 Fiscal Year Final Research Report Summary
Interaction of Bifidobacterium and colonic mucin via carbohydrate-lectin interaction
| Project/Area Number |
06672170
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Biological pharmacy
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| Research Institution | The University of Tokyo |
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Principal Investigator |
YAMAMOTO Kazuo The University of Tokyo, Faculty of Pharmaceutical Sciences, Assistant Professor, 薬学部, 助手 (20174782)
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| Project Period (FY) |
1994 – 1995
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| Keywords | lectin / Bifidobacterium / mucin |
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| Research Abstract |
To determine the specific interactions between Bifidobacterium and colonic mucin, I focused on the interaction of carbohydrate-binding protein (s) expressed on the bacterial cells and colonal mucins. I scanned the ability of B.adolescentis strain Ba56, which were normal inhabitant of the human adult colon, to adhere to the immobilized mucosal components. B.adlescentis was shown to adhere to sulfated mucin, which is expressed on the normal colon. The binding of bacterial cells to sulfated mucin was inhibited by galactose and factose. From the cell surface components of B.adlescentis cells, 44kDa and 30kDa proteins were purified by using affinity chromatography on a column of galactose-Sepharose 4B.The purfied 44 kDa and 30 kDa proteins specifically bound to sulfated mucin expressed on LS174T cells. Benzyl-GalNAc treatment or anti-sulfated mucin monoclonal antibody 91.9H treatment of LS174T cells abolished the binding of ^<125>I-labeled 44kDa and 30kDa proteins. Purified 44 kDa was digested with endoproteinase Lys-C and its proteolytic fragments were recovered by reversed phase HPLC.Partial amino acid sequences of these fragments were analyzed by use of gas-phase protein sequencer.
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