Abstract
The paper describes the reactivity of fluorescein isothiocyanate towards the N-terminus of cytochromes P450 2B4 and 1A2 in solution, in the natural membrane of microsomes and in proteoliposomes (cholate and ultrasonic). The results obtained indicate, that the N-terminus of microsomal or proteoliposomal cytochromes P-450 2B4 and 1A2 spans the membrane only once and faces the vesicles interior. It was suggested that of major importance in orientation of N-terminal residues in the membrane is not the hydrophobic segment itself but rather the positively charged fragment, following it.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Animals
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Aryl Hydrocarbon Hydroxylases*
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Cytochrome P-450 CYP1A2
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Cytochrome P-450 Enzyme System / chemistry
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Cytochrome P-450 Enzyme System / isolation & purification
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Cytochrome P-450 Enzyme System / metabolism*
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Electrophoresis, Polyacrylamide Gel
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Fluorescein-5-isothiocyanate / chemistry*
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Microsomes / drug effects
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Microsomes / enzymology*
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Microsomes / ultrastructure
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Molecular Sequence Data
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Octoxynol / pharmacology
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Oxidoreductases / chemistry
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Oxidoreductases / isolation & purification
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Oxidoreductases / metabolism*
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Proteolipids / metabolism
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Rabbits
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Spectrometry, Fluorescence
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Steroid Hydroxylases / chemistry
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Steroid Hydroxylases / isolation & purification
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Steroid Hydroxylases / metabolism*
Substances
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Proteolipids
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proteoliposomes
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Octoxynol
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Cytochrome P-450 Enzyme System
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Oxidoreductases
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Steroid Hydroxylases
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Aryl Hydrocarbon Hydroxylases
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Cytochrome P-450 CYP1A2
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steroid 15-alpha-hydroxylase
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Fluorescein-5-isothiocyanate