Document Detail

Successful prediction of substrate-binding pocket in SLC17 transporter sialin.
MedLine Citation:
PMID:  22334707     Owner:  NLM     Status:  MEDLINE    
Secondary active transporters from the SLC17 protein family are required for excitatory and purinergic synaptic transmission, sialic acid metabolism, and renal function, and several members are associated with inherited neurological or metabolic diseases. However, molecular tools to investigate their function or correct their genetic defects are limited or absent. Using structure-activity, homology modeling, molecular docking, and mutagenesis studies, we have located the substrate-binding site of sialin (SLC17A5), a lysosomal sialic acid exporter also recently implicated in exocytotic release of aspartate. Human sialin is defective in two inherited sialic acid storage diseases and is responsible for metabolic incorporation of the dietary nonhuman sialic acid N-glycolylneuraminic acid. We built cytosol-open and lumen-open three-dimensional models of sialin based on weak, but significant, sequence similarity with the glycerol-3-phosphate and fucose permeases from Escherichia coli, respectively. Molecular docking of 31 synthetic sialic acid analogues to both models was consistent with inhibition studies. Narrowing the sialic acid-binding site in the cytosol-open state by two phenylalanine to tyrosine mutations abrogated recognition of the most active analogue without impairing neuraminic acid transport. Moreover, a pilot virtual high-throughput screening of the cytosol-open model could identify a pseudopeptide competitive inhibitor showing >100-fold higher affinity than the natural substrate. This validated model of human sialin and sialin-guided models of other SLC17 transporters should pave the way for the identification of inhibitors, glycoengineering tools, pharmacological chaperones, and fluorescent false neurotransmitters targeted to these proteins.
Nicolas Pietrancosta; Christine Anne; Horst Prescher; Raquel Ruivo; Corinne Sagné; Cécile Debacker; Hugues-Olivier Bertrand; Reinhard Brossmer; Francine Acher; Bruno Gasnier
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2012-02-13
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  287     ISSN:  1083-351X     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  2012 Mar 
Date Detail:
Created Date:  2012-07-03     Completed Date:  2012-09-20     Revised Date:  2013-06-26    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  11489-97     Citation Subset:  IM    
Centre National de la Recherche Scientifique, UMR 8601, Université Paris Descartes, Sorbonne Paris Cité, 45 rue des Saints-Pères, F-75006 Paris, France.
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MeSH Terms
Azepines / metabolism
Binding Sites
Computational Biology*
Drug Evaluation, Preclinical
HEK293 Cells
Indoles / metabolism
Models, Molecular
Mutagenesis, Site-Directed
Mutation, Missense
Organic Anion Transporters / chemistry*,  genetics,  metabolism*
Pilot Projects
Protein Binding
Protein Conformation
Sequence Homology, Amino Acid
Sialic Acids / chemistry,  metabolism
Structure-Activity Relationship
Symporters / chemistry*,  genetics,  metabolism*
Reg. No./Substance:
0/Azepines; 0/Indoles; 0/Organic Anion Transporters; 0/Sialic Acids; 0/Symporters; 0/sialic acid transport proteins; 142375-60-8/FR 139317

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