NDLI: Probing protein heterogeneity in the plasma membrane using PALM and pair correlation analysis.

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Probing protein heterogeneity in the plasma membrane using PALM and pair correlation analysis.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Sengupta, Prabuddha Jovanovic-Talisman, Tijana Skoko, Dunja Renz, Malte Veatch, Sarah L. Lippincott-Schwartz, Jennifer
Description	Country affiliation: United States Author Affiliation: Sengupta P ( The Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland, USA.)
Abstract	Photoactivated localization microscopy (PALM) is a powerful approach for investigating protein organization, yet tools for quantitative, spatial analysis of PALM datasets are largely missing. Combining pair-correlation analysis with PALM (PC-PALM), we provide a method to analyze complex patterns of protein organization across the plasma membrane without determination of absolute protein numbers. The approach uses an algorithm to distinguish a single protein with multiple appearances from clusters of proteins. This enables quantification of different parameters of spatial organization, including the presence of protein clusters, their size, density and abundance in the plasma membrane. Using this method, we demonstrate distinct nanoscale organization of plasma-membrane proteins with different membrane anchoring and lipid partitioning characteristics in COS-7 cells, and show dramatic changes in glycosylphosphatidylinositol (GPI)-anchored protein arrangement under varying perturbations. PC-PALM is thus an effective tool with broad applicability for analysis of protein heterogeneity and function, adaptable to other single-molecule strategies.
File Format	HTM / HTML
ISSN	15487091
e-ISSN	15487105
DOI	10.1038/nmeth.1704
Journal	Nature Methods
Issue Number	11
Volume Number	8
Language	English
Publisher	Nature Publishing Group
Publisher Date	2011-09-18
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Clinical Laboratory Techniques Membrane Proteins Metabolism Algorithms Research Support, N.i.h., Extramural Validation Studies
Content Type	Text
Resource Type	Article
Subject	Cell Biology Biochemistry Molecular Biology Biotechnology

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4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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