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  • Reversible binding and rapid diffusion of proteins in complex with inositol lipids serves to coordinate free movement with spatial information.

Reversible binding and rapid diffusion of proteins in complex with inositol lipids serves to coordinate free movement with spatial information.

The Journal of cell biology (2009-01-21)
Gerald R V Hammond, Yirong Sim, Leon Lagnado, Robin F Irvine
ABSTRACT

Polyphosphoinositol lipids convey spatial information partly by their interactions with cellular proteins within defined domains. However, these interactions are prevented when the lipids' head groups are masked by the recruitment of cytosolic effector proteins, whereas these effectors must also have sufficient mobility to maximize functional interactions. To investigate quantitatively how these conflicting functional needs are optimized, we used different fluorescence recovery after photobleaching techniques to investigate inositol lipid-effector protein kinetics in terms of the real-time dissociation from, and diffusion within, the plasma membrane. We find that the protein-lipid complexes retain a relatively rapid ( approximately 0.1-1 microm(2)/s) diffusion coefficient in the membrane, likely dominated by protein-protein interactions, but the limited time scale (seconds) of these complexes, dictated principally by lipid-protein interactions, limits their range of action to a few microns. Moreover, our data reveal that GAP1(IP4BP), a protein that binds PtdIns(4,5)P(2) and PtdIns(3,4,5)P(3) in vitro with similar affinity, is able to "read" PtdIns(3,4,5)P(3) signals in terms of an elongated residence time at the membrane.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Insulin human, meets USP testing specifications
Sigma-Aldrich
Insulin human, ≥95% (HPLC), semisynthetic, powder, non-sterile
Sigma-Aldrich
Insulin human, recombinant, expressed in yeast (proprietary host)
Sigma-Aldrich
Insulin human, recombinant, expressed in yeast, γ-irradiated, suitable for cell culture