PMID: 2502182Jul 6, 1989Paper

Selective chemical modification of cytochrome P-450SCC lysine residues. Identification of lysines involved in the interaction with adrenodoxin

Biochimica Et Biophysica Acta
T B AdamovichS A Usanov

Abstract

Selective chemical modification of cytochrome P-450SCC has been carried out with lysine-modifying reagents. Modification of cytochrome P-450SCC with succinic anhydride was shown to result in loss of its ability to interact with intermediate electron transfer protein - adrenodoxin. To identify amino acid residues involved in charge-ion pairing with complementary carboxyl groups of adrenodoxin, cytochrome P-450SCC complex with adrenodoxin was modified with succinic anhydride. Adrenodoxin was then removed and cytochrome P-450 was additionally modified with isotopically labelled reagent. Subsequent chymotryptic hydrolysis of [14C]succinylated cytochrome P-450SCC and separation of digest obtained by combining various types of HPLC resulted in seven major radioactive peptides. The amino acid sequence of the peptides was determined by microsequencing. The major amino groups modified with radioactive succinic anhydride were found to be at Lys-73, -109, -110, -126, -145, -148 and -154 in the N-terminal sequence of cytochrome P-450SCC molecule and at Lys-267, -270, -338 and -342 in the C-terminal sequence. The role of electrostatic interactions in fixation of cytochrome P-450SCC complex with adrenodoxin is discussed.

References

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Citations

Sep 16, 2005·Journal of Computer-aided Molecular Design·Eszter HazaiDavid Kupfer
Sep 23, 1998·The Journal of Steroid Biochemistry and Molecular Biology·D F Lewis, P Lee-Robichaud
Jun 4, 2011·Proceedings of the National Academy of Sciences of the United States of America·Natallia StrushkevichHee-Won Park
Jan 1, 1993·Drug Metabolism Reviews·L KoymansN P Vermeulen
Jun 1, 1993·DNA and Cell Biology·J A HarikrishnaW L Miller
Jul 19, 2000·Proteins·A V GrinbergR Bernhardt
Jul 22, 1991·Proceedings. Biological Sciences·J E BaldwinW G Richards
Jun 8, 2004·The Journal of Biological Chemistry·Burkhard SchifflerRita Bernhardt
Jan 28, 1991·FEBS Letters·S A Usanov, V L Chashchin
Jul 23, 2003·Chemical Research in Toxicology·Zhongfa LiuLawrence M Sayre
Jan 10, 2014·Chemical Reviews·Thomas L Poulos
Jun 12, 2010·Biochimica Et Biophysica Acta·Kerstin Maria EwenRita Bernhardt

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