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PMID: 16829131 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Review

Determining selectivity of phosphoinositide-binding domains.

Methods (San Diego, Calif.) ·Vol. 39 ·No. 2 ·2006-06-00 ·Pages 122-33

Narayan K, Lemmon MA

Abstract

The burgeoning of phosphoinositide-binding domains and proteins in cellular signaling and trafficking has drawn laboratories from a wide variety of fields into the study of lipid interactions with peripheral membrane proteins. Many different approaches have been developed to assess phosphoinositide binding, some of which are more problematic than others, and some of which can be quantitated more readily than others. With a focus on the methods used in our laboratory, we describe here the considerations that need to be taken into account when establishing-and quantitating-the specific binding of a protein or domain to phosphoinositides in membranes. We also discuss briefly a few examples in which no clear consensus has yet been reached as to the specificity of a given domain or protein because of discrepancies between different commonly used approaches.

MeSH Terms
Binding Sites Biochemistry/methods Blotting, Western Calorimetry/methods Kinetics Lipids/analysis Phosphatidylinositols/metabolism Phospholipids/analysis,metabolism Phosphorus Radioisotopes Protein Structure, Tertiary Proteins/metabolism Substrate Specificity Surface Plasmon Resonance
Chemicals
Lipids Phosphatidylinositols Phospholipids Phosphorus Radioisotopes Proteins
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Narayan Kartik
Department of Biochemistry and Biophysics, University of Pennsylvania School of Medicine, Philadelphia, PA 19104-6059, USA.
Lemmon Mark A
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Article Info
Journal
Methods (San Diego, Calif.)
Abbr.
Methods
ISSN
1046-2023
DOI
10.1016/j.ymeth.2006.05.006
Published
2006-06-00
Pages
122-33
Language
English
Region
United States
NLM ID
9426302
PMCID
PMC3786563
Subset
IM
Grants
NIGMS NIH HHS · R01 GM056846 · United States
NIGMS NIH HHS · F32 GM073496 · United States
NIGMS NIH HHS · R01-GM056846 · United States
NIGMS NIH HHS · R01 GM056846-07 · United States
NIGMS NIH HHS · F32-GM073496 · United States
External Links
PubMed source DOI: 10.1016/j.ymeth.2006.05.006 PMC Full Text View journal details
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