| Budget Amount *help |
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1991: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1990: ¥1,100,000 (Direct Cost: ¥1,100,000)
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| Research Abstract |
Heat shock protein of molecular weight 47, 000 D, HSP47, binds to native and denatured collagen including type I, type III, or type IV. HSP47 is supposed to be implicated in the transportation of procollagen molecules from ER to Golgi. In order to determine immunohistochemically the distribution of HSP47 in liver or lung from the patients with fibrotic diseases, polyclonal and monoclonal antibodies against chicken HSP47 were used. Howerer, in the fixed mammalian tissues such antidobies were not able to atain HSP47 because of the disruption of the epitopes by fixation. Therefore, in order to get the antibody that react to human HSP47 even in the fixed tissues, human HSP47 was partially purified from NP-40 extract of human embryonal fibroblasts cultured in at least 10 roller bottles by gelatin affinity chromatography or immunoaffinity chromatography. It was very difficult to get further purified HSP47 because human HSP47 in the partially purified fraction was extremely unstable and easily degraded. This difficulty disturbed the production of the specific polyclonal or monoclonal antibody against human HSP47. Thus, immunohistochemical study for the distribution of HSP47 in the fibrotic diseases was not proceeded.
The trial of in situ hybridization with cDNA probe of mouse HSP47 in the section of liver or lung tissue of the fibrotic diseases failed to determine the distribution of mRNA for HSP47.
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