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Molecular Analysis of Carotenoid Cyclase Inhibition
- Source :
- Archives of Biochemistry and Biophysics. 346:53-64
- Publication Year :
- 1997
- Publisher :
- Elsevier BV, 1997.
-
Abstract
- Later steps of carotenoid biosynthesis catalyzed by cyclase enzymes involve the formation of alpha, beta, and kappa-rings. Examination of the primary structure of lycopene beta-cyclase revealed 55% identity with that of antheraxanthin kappa-cyclase. Recombinant lycopene beta-cyclase afforded only beta-carotene, while recombinant antheraxanthin kappa-cyclase catalyzed the formation of beta-carotene from lycopene as well as the conversion of antheraxanthin into the kappa-carotenoid capsanthin. Since the formation of beta- and kappa-rings involves a transient carotenoid carbocation, this suggests that both cyclases initiate and/or neutralize the incipient carbocation by similar mechanisms. Several amine derivatives protonated at physiological pH were used to examine the molecular basis of this phenomenon. The beta-and kappa-cyclases displayed similar inhibition patterns. Affinity or photoaffinity labeling using p-dimethylamino-benzenediazonium fluoroborate, N,N-dimethyl-2-phenylaziridinium, and nicotine irreversibly inactivated both cyclase enzymes. Photoaffinity labeling using [3H]nicotine followed by radiosequence analysis and site-directed mutagenesis revealed the existence of two cyclase domains characterized by the presence of reactive aromatic and carboxylic amino acid residues. We propose that these residues represent the "negative point charges" involved in the coordination of the incipient carotenoid carbocations.
- Subjects :
- Nicotine
Aziridines
Molecular Sequence Data
Biophysics
Carbocation
Biochemistry
Cyclase
chemistry.chemical_compound
Ethylamines
Amino Acid Sequence
Amines
Intramolecular Lyases
Molecular Biology
Carotenoid
Plant Proteins
chemistry.chemical_classification
Binding Sites
Sequence Homology, Amino Acid
Photoaffinity labeling
Antheraxanthin
Mutagenesis
Affinity Labels
Diazonium Compounds
Carotenoids
Lycopene
Enzyme
chemistry
Mutagenesis, Site-Directed
Oxidoreductases
Dimethylamines
Subjects
Details
- ISSN :
- 00039861
- Volume :
- 346
- Database :
- OpenAIRE
- Journal :
- Archives of Biochemistry and Biophysics
- Accession number :
- edsair.doi.dedup.....14e83bdb0afe958be1626be13210c6ad