title: Elucidating the role of select cytoplasmic proteins in altering diffusion of integrin receptors.
abstract: Cytoplasmic proteins that affect integrin diffusion in the cell membrane are identified using a combination of fluorescence recovery after photobleaching (FRAP) and RNA interference. Integrin receptors are essential for many cellular events, and alterations in lateral diffusion are one mechanism for modulating their function. In cells expressing native cytoplasmic protein concentrations and spread on a slide containing integrin extracellular ligand, 45 ± 2% of the integrin is mobile with a time-dependent 5.2 ± 0.9 × 10(-9) cm(2)/s diffusion coefficient at 1 s. The time exponent is 0.90 ± 0.07, indicating integrin diffusion moderately slows at longer times. The role of a specific cytoplasmic protein in altering integrin diffusion is revealed through changes in the FRAP curve after reducing the cytoplasmic protein's expression. Decreased expression of cytoplasmic proteins rhea, focal adhesion kinase (FAK), or steamer duck decreases the integrin mobile fraction. For rhea and FAK, there is a concomitant shift to Brownian (i.e., time-independent) diffusion at reduced concentrations of these proteins. In contrast, when the expression of actin 42A, dreadlocks, paxillin, integrin-linked kinase (ILK), or vinculin is reduced, integrin diffusion generally becomes more constrained with an increase in the integrin mobile fraction. This same change in integrin diffusion is measured in the absence of integrin extracellular ligand. The results indicate breaking the extracellular ligand-integrin-cytoskeletal linkage alters integrin diffusion properties, and, in most cases, there is no correlation between integrin and lipid diffusion properties.
authors.affiliation: Department of Chemistry, Iowa State University, Ames, IA 50011, USA.
authors.forename: Suzanne
authors.initials: S
authors.lastname: Sander
authors.forename: Neha
authors.initials: N
authors.lastname: Arora
authors.forename: Emily A
authors.initials: EA
authors.lastname: Smith
mesh_terms: D000818:Animals; D002460:Cell Line; D003598:Cytoskeletal Proteins / Q000235:genetics* / Q000378:metabolism*; D004058:Diffusion; D036681:Fluorescence Recovery After Photobleaching / Q000379:methods; D016023:Integrins / Q000378:metabolism*; D008563:Membrane Lipids / Q000378:metabolism; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics
chemical_list: D003598:Cytoskeletal Proteins; D016023:Integrins; D008563:Membrane Lipids; D012333:RNA, Messenger