-transparent.png){kind=logo}
Three fractions of cytochrome P-450scc (denoted as fractions a, b, and c) were purified by a new procedure from bovine adrenocortical mitochondria. The amino-acid content analyses of these three fractions showed no difference. NH2-terminal amino-acid sequences of cytochrome P-450scc fractions, a and b agreed completely with the sequence deduced by nucleotide sequence of cDNA of cytochrome P-450scc mRNA (Morohashi, K., Fujii-Kuriyama, Y., Okada, Y., Sogawa, K., Hirose, T., Inayama, S. and Omura, T. (1984) Proc. Natl. Acad. Sci. USA 81, 4647-4651), whereas the sequence of fraction c showed a missing of isoleucine at the NH2-terminal. COOH-terminal ámino-acid sequences of fractions a, b and c were -Gln-Ala-COOH, identical with the deduced sequence from the cDNA. Measurements of the enzymatic activities of cholesterol side-chain cleavage reaction revealed no distinct difference among these three fractions. Although each of these fractions appeared as a single protein staining band upon SDS-polyacrylamide gel electrophoresis, these fractions showed heterogeneities upon two-dimensional electrophoresis and chromatofocusing. Fraction a contained the major form of cytochrome P-450scc, and its isoelectric point was estimated to be pH 7.8 by isoelectric focusing under both native and denatured conditions, and this value was confirmed by chromatofocusing. Neither of the carbohydrate-specific stainings (such as periodic acid-Schiff staining and lectin-peroxidase stainings using concanavalin A, wheat-germ agglutinin, and soybean agglutinin) of purified cytochrome P-450scc fractions after the electrophoretic resolution on SDS-polyacrylamide gel could show cytochrome P-450scc fractions as glycoproteins, suggesting that the heterogeneities were not due to the glycosylation state.
M Tsubaki, and
H Ohkubo, and
Y Tsuneoka, and
S Tomita, and
A Hiwatashi, and
Y Ichikawa
February 1989,
Biochimica et biophysica acta,
M Tsubaki, and
H Ohkubo, and
Y Tsuneoka, and
S Tomita, and
A Hiwatashi, and
Y Ichikawa
June 1988,
FEBS letters,
M Tsubaki, and
H Ohkubo, and
Y Tsuneoka, and
S Tomita, and
A Hiwatashi, and
Y Ichikawa
January 1976,
Advances in experimental medicine and biology,
M Tsubaki, and
H Ohkubo, and
Y Tsuneoka, and
S Tomita, and
A Hiwatashi, and
Y Ichikawa
March 1979,
Biochimica et biophysica acta,
M Tsubaki, and
H Ohkubo, and
Y Tsuneoka, and
S Tomita, and
A Hiwatashi, and
Y Ichikawa
May 1993,
Biochimica et biophysica acta,
M Tsubaki, and
H Ohkubo, and
Y Tsuneoka, and
S Tomita, and
A Hiwatashi, and
Y Ichikawa
February 1981,
Biochemistry,
M Tsubaki, and
H Ohkubo, and
Y Tsuneoka, and
S Tomita, and
A Hiwatashi, and
Y Ichikawa
June 1991,
The Journal of steroid biochemistry and molecular biology,
M Tsubaki, and
H Ohkubo, and
Y Tsuneoka, and
S Tomita, and
A Hiwatashi, and
Y Ichikawa
June 1984,
Biochimica et biophysica acta,
M Tsubaki, and
H Ohkubo, and
Y Tsuneoka, and
S Tomita, and
A Hiwatashi, and
Y Ichikawa
January 1991,
The International journal of biochemistry,
M Tsubaki, and
H Ohkubo, and
Y Tsuneoka, and
S Tomita, and
A Hiwatashi, and
Y Ichikawa
October 1980,
European journal of biochemistry,
