roshni
04-04-2006, 06:29 PM
Biophys J. (javascript:AL_get(this, 'jour', 'Biophys J.');) 2006 Mar 31; [Epub ahead of print] (http://www.ncbi.nlm.nih.gov/entrez/utils/lofref.fcgi?PrId=3051&uid=16581836&db=pubmed&url=http://www.biophysj.org/cgi/pmidlookup?view=long&pmid=16581836)
Quantification of {alpha}-Synuclein binding to lipid vesicles using fluorescence correlation spectroscopy.
Rhoades E (http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Search&itool=pubmed_Abstract&term=%22Rhoades+E%22%5BAuthor%5D), Ramlall TF (http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Search&itool=pubmed_Abstract&term=%22Ramlall+TF%22%5BAuthor%5D), Webb WW (http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Search&itool=pubmed_Abstract&term=%22Webb+WW%22%5BAuthor%5D), Eliezer D (http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Search&itool=pubmed_Abstract&term=%22Eliezer+D%22%5BAuthor%5D).
Cornell University.
alpha-Synuclein (alphaS) is a soluble synaptic protein that is the major proteinaceous component of insoluble fibrillar Lewy body deposits that are the hallmark of Parkinson's disease (PD). The interaction of alphaS with synaptic vesicles is thought to be critical both to its normal function as well as to its pathological role in PD. We demonstrate the use of fluorescence correlation spectroscopy (FCS) as a tool for rapid and quantitative analysis of the binding of alphaS to large unilamellar vesicles of various lipid compositions. We find that alphaS binds preferentially to vesicles containing acidic lipids, and that this interaction can be blocked by increasing the concentration of NaCl in solution. Negative charge is not the only factor determining binding, as we clearly observe binding to vesicles composed entirely of zwitterionic lipids. Additionally, we find enhanced binding to lipids with less bulky head groups. Quantification of the protein to lipid ratio required for binding to different lipid compositions, combined with other data in the literature, yields an upper bound estimate for the number of lipid molecules required to bind each individual molecule of alphaS. Our results demonstrate that FCS provides a powerful tool for the quantitative characterization of alphaS-lipid interactions.
PMID: 16581836 [PubMed - as supplied by publisher]
Quantification of {alpha}-Synuclein binding to lipid vesicles using fluorescence correlation spectroscopy.
Rhoades E (http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Search&itool=pubmed_Abstract&term=%22Rhoades+E%22%5BAuthor%5D), Ramlall TF (http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Search&itool=pubmed_Abstract&term=%22Ramlall+TF%22%5BAuthor%5D), Webb WW (http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Search&itool=pubmed_Abstract&term=%22Webb+WW%22%5BAuthor%5D), Eliezer D (http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Search&itool=pubmed_Abstract&term=%22Eliezer+D%22%5BAuthor%5D).
Cornell University.
alpha-Synuclein (alphaS) is a soluble synaptic protein that is the major proteinaceous component of insoluble fibrillar Lewy body deposits that are the hallmark of Parkinson's disease (PD). The interaction of alphaS with synaptic vesicles is thought to be critical both to its normal function as well as to its pathological role in PD. We demonstrate the use of fluorescence correlation spectroscopy (FCS) as a tool for rapid and quantitative analysis of the binding of alphaS to large unilamellar vesicles of various lipid compositions. We find that alphaS binds preferentially to vesicles containing acidic lipids, and that this interaction can be blocked by increasing the concentration of NaCl in solution. Negative charge is not the only factor determining binding, as we clearly observe binding to vesicles composed entirely of zwitterionic lipids. Additionally, we find enhanced binding to lipids with less bulky head groups. Quantification of the protein to lipid ratio required for binding to different lipid compositions, combined with other data in the literature, yields an upper bound estimate for the number of lipid molecules required to bind each individual molecule of alphaS. Our results demonstrate that FCS provides a powerful tool for the quantitative characterization of alphaS-lipid interactions.
PMID: 16581836 [PubMed - as supplied by publisher]