Visualization and manipulation of phosphoinositide dynamics in live cells using engineered protein domains.
There is hardly a membrane-associated molecular event that is not regulated by phosphoinositides, a minor but critically important class of phospholipids of cellular membranes. The rapid formation, elimination, and conversion of these lipids in specific membrane compartments are ensured by a wealthy number of inositol lipid kinases and phosphatases with unique localization and regulatory properties. The existence of multiple inositol lipid pools have been indicated by metabolic labeling studies, but the level of functional compartmentalization revealed by the identification of numerous protein effectors acted upon by phosphoinositides could not have been foreseen. The changing perception of inositides from just serving as lipid precursors of second messengers to becoming highly dynamic local membrane-bound regulators poses new challenges concerning the detection of their rapid localized changes. Moreover, it is increasingly evident that manipulation of lipids in highly defined compartments would be a highly superior approach to soaking the cells with a particular phosphoinositide when studying the local regulation of the lipid on any effectors. In this review, we will summarize our efforts to improve our tools in studying phosphoinositide dynamics and discuss our views on the values of these methods compared to other options currently used or being explored.
Várnai
Péter
P
Endocrinology and Reproduction Research Branch, NICHD, National Institutes of Health, Bldg 49, Rm 6A35, 49 Convent Drive, Bethesda, MD, USA. varnai@puskin.sote.hu
Balla
Tamas
T
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D002477:Cells / Q000378:metabolism* / Q000648:ultrastructure*; D006801:Humans; D007295:Inositol Phosphates / Q000378:metabolism; D018613:Microscopy, Confocal; D010716:Phosphatidylinositols / Q000378:metabolism*; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000494:pharmacology
D007295:Inositol Phosphates; D010716:Phosphatidylinositols; D011994:Recombinant Proteins