PMID:8341666
| Citation |
Barker, PD, Ferrer, JC, Mylrajan, M, Loehr, TM, Feng, R, Konishi, Y, Funk, WD, MacGillivray, RT and Mauk, AG (1993) Transmutation of a heme protein. Proc. Natl. Acad. Sci. U.S.A. 90:6542-6 |
|---|---|
| Abstract |
Residue Asn57 of bovine liver cytochrome b5 has been replaced with a cysteine residue, and the resulting variant has been isolated from recombinant Escherichia coli as a mixture of four major species: A, BI, BII, and C. A combination of electronic spectroscopy, 1H NMR spectroscopy, resonance Raman spectroscopy, electrospray mass spectrometry, and direct electrochemistry has been used to characterize these four major cytochrome derivatives. The red form A (E(m) = -19 mV) is found to possess a heme group bound covalently through a thioether linkage involving Cys57 and the alpha carbon of the heme 4-vinyl group. Form BI has a covalently bound heme group coupled through a thioether linkage involving the beta carbon of the heme 4-vinyl group. Form BII is similar to BI except that the sulfur involved in the thioether linkage is oxidized to a sulfoxide. The green form C (E(m) = 175 mV) possesses a noncovalently bound prosthetic group with spectroscopic properties characteristic of a chlorin. A mechanism is proposed for the generation of these derivatives, and the implications of these observations for the biosynthesis of cytochrome c and naturally occurring chlorin prosthetic groups are discussed. |
| Links | |
| Keywords |
Animals; Cattle; Cysteine/chemistry; Cysteine/genetics; Cytochromes b5/chemistry; Cytochromes b5/genetics; Genetic Variation; Magnetic Resonance Spectroscopy; Mutagenesis, Site-Directed; Mutation; Peptide Fragments/chemistry; Peptide Fragments/genetics; Protoporphyrins/chemistry; Recombinant Proteins/chemistry; Recombinant Proteins/genetics; Spectrophotometry; Spectrum Analysis, Raman |
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