Document Detail


Ascorbic acid-induced crosslinking of lens proteins: evidence supporting a Maillard reaction.
MedLine Citation:
PMID:  3408736     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The incubation of calf lens extracts with 20 mM ascorbic acid under sterile conditions for 8 weeks caused extensive protein crosslinking, which was not observed with either 20 mM sorbitol or 20 mM glucose. While no precipitation was observed, ascorbic acid did induce the formation of high-molecular-weight protein aggregates as determined by Agarose A-5m chromatography. Proteins modified by ascorbic acid bound strongly to a boronate affinity column, however, crosslinked proteins were present mainly in the unbound fraction. These observations suggest that the cis-diol groups of ascorbic acid were present in the primary adduct, but were either lost during the crosslinking reaction or sterically hindered from binding to the column matrix. The amino acid composition of the ascorbic acid-modified proteins was identical to controls except for a 15% decrease in lysine. Amino acid analysis after borohydride reduction, however, showed a 25% decrease in lysine, a 7% decrease in arginine and an additional peak which eluted between phenylalanine and histidine. Extensive browning occurred during the ascorbic acid-modification reaction. This resulted in protein-bound chromophores with a broad absorption spectrum from 300 to 400 nm, and protein-bound fluorophores with excitation/emission maxima of 350/450 nm. A 4 week incubation of dialyzed crude lens extract with [1-14C]ascorbic acid showed increased incorporation for 2 weeks, followed by a decrease over the next 2 weeks as crosslinking was initiated. The addition of cyanoborohydride to the reaction mixture completely inhibited crosslinking and increased [1-14C]ascorbic acid incorporation to a plateau value of 180 nmol per mg protein. Amino acid analysis showed a 50% loss of lysine, and 8% decrease in arginine and the presence of a new peak which eluted slightly earlier than methionine. These data are consistent with the non-enzymatic glycation of lens proteins by either ascorbic acid or an oxidation product of ascorbic acid via a Maillard-type reaction.
Authors:
B J Ortwerth; P R Olesen
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Publication Detail:
Type:  In Vitro; Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Biochimica et biophysica acta     Volume:  956     ISSN:  0006-3002     ISO Abbreviation:  Biochim. Biophys. Acta     Publication Date:  1988 Aug 
Date Detail:
Created Date:  1988-10-07     Completed Date:  1988-10-07     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0217513     Medline TA:  Biochim Biophys Acta     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  10-22     Citation Subset:  IM    
Affiliation:
Mason Institute of Ophthalmology, University of Missouri, Columbia 65212.
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MeSH Terms
Descriptor/Qualifier:
Amino Acids / analysis
Animals
Ascorbic Acid*
Borohydrides
Cattle
Cross-Linking Reagents*
Crystallins*
Lens, Crystalline / physiology*
Molecular Weight
Oxidation-Reduction
Spectrophotometry, Ultraviolet
Grant Support
ID/Acronym/Agency:
EY 02035/EY/NEI NIH HHS; EY 07070/EY/NEI NIH HHS
Chemical
Reg. No./Substance:
0/Amino Acids; 0/Borohydrides; 0/Cross-Linking Reagents; 0/Crystallins; 50-81-7/Ascorbic Acid

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


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