Document Detail


Purification and partial characterization of a cellular carotenoid-binding protein from ferret liver.
MedLine Citation:
PMID:  9305906     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A cellular carotenoid-binding protein was purified to homogeneity from beta-carotene-fed ferret liver utilizing the following steps: ammonium sulfate precipitation, ion exchange, gel filtration, and affinity chromatography. The final purification was 607-fold. [14C]beta-Carotene co-purified with the binding protein throughout the purification procedures. SDS-PAGE of the purified protein showed a single band with an apparent molecular mass of 67 kDa. Scatchard analysis of the specific binding of the purified protein to beta-carotene showed two classes of binding sites, a high affinity site with an apparent Kd of 56 x 10(-9) M and a low affinity site with a Kd of 32 x 10(-6) M. The Bmax for beta-carotene binding to the high affinity site was 1 mol/mol, while that for the low affinity site was 145 mol/mol. The absorption spectrum of the complex showed a 32-nm bathochromic shift in lambdamax with minor peaks at 460 and 516 nm. Except for alpha-carotene and cryptoxanthin, none of the model carotenoids or retinol competed with beta-carotene binding to the protein. Thus, a specific carotenoid-binding protein of 67 kDa has been characterized in mammalian liver with a high degree of specificity for binding only carotenoids with at least one unsubstituted beta-ionone ring.
Authors:
M N Rao; P Ghosh; M R Lakshman
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  272     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  1997 Sep 
Date Detail:
Created Date:  1997-10-23     Completed Date:  1997-10-23     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  24455-60     Citation Subset:  IM    
Affiliation:
Department of Medicine, George Washington University, Washington, D. C. 20037, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Binding, Competitive
Chromatography, Affinity
Chromatography, Gel
Chromatography, Ion Exchange
Electrophoresis, Polyacrylamide Gel
Ferrets
Ligands
Liver / metabolism*
Male
Proteins / isolation & purification*,  metabolism
beta Carotene / administration & dosage,  metabolism*
Grant Support
ID/Acronym/Agency:
CA39999/CA/NCI NIH HHS
Chemical
Reg. No./Substance:
0/Ligands; 0/Proteins; 7235-40-7/beta Carotene

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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