2000 Fiscal Year Final Research Report Summary
Molecular and biological studies of colonization factors of oral spirochetes
| Project/Area Number |
11671830
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Morphological basic dentistry
|
|---|
| Research Institution | Asahi University |
|---|
Principal Investigator |
UMEMOTO Toshihiko School of Dentistry, Department of Oral Microbiology, Asahi University Associate Professor, 歯学部・口腔細菌学講座, 助教授 (80076033)
|
|---|
| Project Period (FY) |
1999 – 2000
|
| Keywords | oral spirochetes / colonization factors / coaggregation / chemotaxis / albumin-binding proteins / flagellar genes |
|---|
| Research Abstract |
The aim of this poject was to investigate molecular and biological analyses of colonization factors of human oral spirochetes. The Porphyromonas gingivalis fimbria-binding polypeptide, fragellar genes, and albumin-binding polypeptides of a medium-sized Treponema medium ATCC 700293 were focused.
1. Immunoelectron microscopy demonstrated that the 37 kDa fimbria-binding polypeptide was localized on the outer envelope. The surface protein was partially purified from SDS solubilized cells by the use of several chromatography matrices, including affinity chromatography on a polyclonal anti-37 kDa polypeptide serum-CNBr-activated Sepahrose 4B matrix. The purified protein had an IpH of 7.4, and exhibited the property of binding to the fimbria from P.gingivalis ATCC 33277 and 381.
2. The DNA fragment of the T.medium was cloned and sequenced. The size of the fragment was 15,809 bp including flagellar genes flgD and flgE.The flgD sequence was 480 bp in length, and flgE sequence (873 bp) was separat
… More
ed with a transposon-like sequence.. The nucleotide sequence of T.medium flgD and flgE genes were relatively preserved when compared with other host-associated treponemal species.
3. The motility and chemotaxis of human oral spirochetes Treponema denticola ATCC 35404, T.medium, and T.vincentii ATCC 35580 were examined by a capillary assay method. Of five tested sera three human oral treponemes were chemoattractant to rabbit serum. The checkerboard analysis of chemotaxis toward the rabbti serum showed that T.denticola cells swam toward the culutre media containing higher concentrations of the serum. The treponemal chemotaxis was reduced by heating serum, and increased by addition of rabbit albumin. Western blotting analysis demonstrated that these treponemes possessed rabbit albumin-binding polypeptides with molecular sizes of 65 kDa and 70 kDa. Immunoelectron microscopy demonstrated that the 65 kDa rabbit albumin-binding polypeptide was located on their outer envelopes, suggesting that the rabbit albumin-binding polypeptide seem to be responsible for chemotaxis toward rabbit serum. Less
|
Research Products
(2 results)
