Distribution of the mitogenic protein, sclerogen, in extracts from Sclerotinia sclerotiorum IFO 9395 assessed by using immunochemical analysis.
Sclerogen is a mitogenic protein isolated from the buffer extracts (3S) from sclerotia and shows significant mitogenic activity after heat denaturation accompanied with conformational changes. Experiments have been carried out to assess the distribution of sclerogen in several extracts from sclerotia and mycelia of this fungus by using the anti-serum against sclerogen. The anti-serum against the native- and heat denatured-sclerogen (anti-sclerogen-M and -MB serum, respectively) were prepared from Japanese white rabbits. These anti-sera showed different cross-reactivity to sclerogen-M and -MB from the results of immunoblotting after normal-polyacrylamide gel electrophoresis (normal-PAGE), suggesting that sclerogen-MB became to have different epitope(s) after heat denaturation of sclerogen-M. The result of immunoblotting after sodium dodecyl sulfate (SDS)-PAGE suggested that these anti-sera also recognized the random coil site(s) where the conformational specificity was not concerned, and that sclerogen (like substance(s] existed not only in mild extracts (e.g. phosphate buffer) but also in fractions extracted from mycelia of S. sclerotiorum IFO 9395. Taken together with the results of the mitogenic activity of the representative extracts from sclerotia, at least two kinds of the mitogenic substances would exist in sclerotia; one would be extracted by mild conditions, and the other would be difficult to extract and require drastic conditions such as hot water extraction.
Tokyo College of Pharmacy, Japan.
H
H
Shinohara
N
N
Ohno
T
T
Yadomae
D000818:Animals; D000918:Antibody Specificity; D001203:Ascomycota / Q000737:chemistry*; D015153:Blotting, Western; D003429:Cross Reactions; D004586:Electrophoresis; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005656:Fungal Proteins / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D008813:Mice, Inbred ICR; D008934:Mitogens / Q000032:analysis*; D008970:Molecular Weight; D011489:Protein Denaturation; D011506:Proteins / Q000032:analysis*
D005656:Fungal Proteins; D008934:Mitogens; D011506:Proteins; C013247:sclerotin