SEQUENCE DETERMINATION OF THE PSEUDOMONAS PUTIDA CYTOCHROME P-450: ISOLATION AND PARTIAL SEQUENCES OF THE CYSTEINE PEPTIDES

Abstract

Publisher Summary This chapter focuses on the sequence determination of the Pseudomonas Putida cytochrome P-450. Pseudomonas putida contains a camphor hydroxylating system, which requires NADH-putidaredoxin reductase, putidaredoxin, and cytochrome P-450. The amino acid sequences of the protein components are essential for investigating the structure–function relationships of the individual components and a multienzyme complex for comparing the chemical structures of the individual components from different living species and for the complete structural determination of the protein components by crystal X-ray diffraction studies. For the goal of studying the structure–function relationships of the cysteine residues in cytochrome P-450 by chemical modification experiments, it was necessary to devise a method for isolating the individual cysteine peptides by tryptic hydrolysis of P-450 cam . A method has been devised for isolating 6 tryptic peptides that contain cysteines. The chapter presents the partial or complete sequences of these peptides and discusses the possible enzymatic or structural roles of the cysteine residues.