Size dependence of the translational diffusion of large integral membrane proteins in liquid-crystalline phase lipid bilayers. A study using fluorescence recovery after photobleaching

Vaz, Winchil L.C.; Criado, Manuel; Madeira, Vı́tor M.C.; Schoellmann, Guenther; Jovin, Thomas M.

doi:10.1021/bi00265a034

Cited by 126 publications

(83 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…4, B and D). Within the region of interest, the recovery profile of eCFP vector is nearly 100%, which is consistent with the diffusion profile of small cytoplasmic unbound proteins (42)(43)(44). The recovery profile of eCFP vector is not altered by the presence or absence of YFP-DAT (Fig.…”

Section: Dopamine Transporter and ␣-Synucleinsupporting

confidence: 79%

Dopamine Transporter Activity Is Modulated by α-Synuclein

Butler

Goodwin

Saha

et al. 2015

Journal of Biological Chemistry

View full text Add to dashboard Cite

Background: DAT regulates dopamine neurotransmission in the brain. Results: ␣-Synuclein influences DA efflux and membrane microdomain distribution of DAT. Conclusion: DAT activation recruits ␣-synuclein to the membrane, which in turn influences dopamine neurotransmission. Significance: Understanding the mechanisms associated with ␣-synuclein regulation of DAT may reveal disease-modifying targets for the treatment of pathologies associated with DA dysregulation.

show abstract

Section: Dopamine Transporter and ␣-Synucleinsupporting

confidence: 79%

Dopamine Transporter Activity Is Modulated by α-Synuclein

Butler

Goodwin

Saha

et al. 2015

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…Compared to N-4-nitrobenzo-2-oxa-1,3-diazole phosphatidylethanolamine, it diffuses at a rate near the predicted theoretical limit for a freely diffusing membrane protein (32)(33)(34) . Similar diffusion coefficients have been measured for rhodopsin in amphibian rod outer segment disks (35,36), and for plasma membrane proteins in membranes in which an underlying cytoskeleton may be modified or is separated from the membrane (37)(38)(39)(40)(41)(42) .…”

Section: Rapid Communicationsmentioning

confidence: 77%

A localized surface protein of guinea pig sperm exhibits free diffusion in its domain.

Myles¹,

Primàkoff²,

Koppel³

1984

The Journal of Cell Biology

View full text Add to dashboard Cite

Using the technique of fluorescence redistribution after photobleaching, we are studying the cellular mechanisms involved in localizing surface molecules to particular domains . A number of antigens localized to discrete surface regions have been identified with monoclonal antibodies on guinea pig sperm cells (Primakoff, P., and D . G . Myles, 1983, Dev . Biol ., 98 :417-428) . One of these monoclonal antibodies, PT-1, binds exclusively to the posterior tail region of the sperm cell surface . PT-1 recognizes an integral membrane protein that in complex with n-octyl-ß-D-glucopyranoside has a sedimentation coefficient of 6 .8S in sucrose density gradients . Fluorescence redistribution after photobleaching measurements reveal that within its surface domain the PT-1 antigen diffuses rapidly (D = 2 .5 x 10 -9 Cm 2/s) and completely (>90% recovery after bleaching) . These results rule out for this membrane protein all models that invoke immobilization as a mechanism for maintaining localization . We propose that the mechanism for localization of the PT-1 antigen may be a barrier to diffusion at the domain boundary .The restriction of cell surface molecules into specific domains has been well established for a wide variety of mammalian cell types . These include acetylcholine receptors at the synapse on muscle cells (1), low density lipoprotein and asialoglycoprotein receptors in coated pits of fibroblasts (2) and hepatocytes (3) and more extensive localized domains on the surface of sperm (4, 5), intestinal epithelial cells-(6, 7), and hepatocytes (8, 9). Although topographical localization of various surface receptors has been widely described, little is known concerning the mechanisms that maintain surface protein localizations . Immobilization in the membrane is a potential mechanism for maintaining a non-uniform distribution of surface molecules, and in the case of the acetylcholine receptor, it has been shown that clustered receptors are immobilized (10).In the current study we have addressed the question whether there are mechanisms other than immobilization to localize surface molecules . We have used the guinea pig sperm cell . Previously, the existence of five different antigen domains on the surface of the guinea pig sperm was established with monoclonal antibodies directed to surface antigens. These antigens may be restricted to the anterior head, posterior head, whole head, whole tail, or posterior tail cell surface (11). In some cases, patching of localized sperm surface molecules can be induced, suggesting that these molecules may be mobile in the plane of the membrane (12-15) . However, ability to patch may not be a reliable indicator of mobility of a receptor in the membrane (16-18) and gives no measure of the diffusion rate or percent mobile fraction. Therefore, we RAPID COMMUNICATIONS have directly measured these parameters using fluorescence redistribution after photobleaching of a localized sperm surface antigen that can be patched . This antigen, identified by the monoclonal antibody PT-...

show abstract

“…In Fig. 2, we gather published diffusion coefficients of transmembrane peptides (15) and various proteins (16). The D variations, normalized by the lipid diffusion, show a continuous decrease of mobility, fitted by a simple 1͞R law.…”

Section: Resultsmentioning

confidence: 99%

Lateral mobility of proteins in liquid membranes revisited

Gambin

López-Esparza

Reffay

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

352

377

View full text Add to dashboard Cite

The biological function of transmembrane proteins is closely related to their insertion, which has most often been studied through their lateral mobility. For >30 years, it has been thought that hardly any information on the size of the diffusing object can be extracted from such experiments. Indeed, the hydrodynamic model developed by Saffman and Delbrü ck predicts a weak, logarithmic dependence of the diffusion coefficient D with the radius R of the protein. Despite widespread use, its validity has never been thoroughly investigated. To check this model, we measured the diffusion coefficients of various peptides and transmembrane proteins, incorporated into giant unilamellar vesicles of 1-stearoyl-2-oleoylsn-glycero-3-phosphocholine (SOPC) or in model bilayers of tunable thickness. We show in this work that, for several integral proteins spanning a large range of sizes, the diffusion coefficient is strongly linked to the protein dimensions. A heuristic model results in a Stokes-like expression for D, (D ؔ 1͞R), which fits literature data as well as ours. Diffusion measurement is then a fast and fruitful method; it allows determining the oligomerization degree of proteins or studying lipid-protein and protein-protein interactions within bilayers.bilayers ͉ transmembrane proteins ͉ diffusion ͉ peptides ͉ sponge phase I n the hydrodynamic model of Saffman and Delbrück (1), transmembrane peptides and proteins are described as diffusing in a perfectly continuous medium, ignoring the finite size of the lipids. This model predicts that the diffusion coefficient D of a simple cylinder embedded in a thin sheet of fluid matching its height ( Fig. 1) is given byIn this expression, the adjustable parameters are by order of importance: the thickness h and viscosity m of the liquid membrane, the radius R of the diffusing cylinder, and the viscosity of the surrounding aqueous phase w . This result follows from solving the flow field in the membrane and in the surrounding fluid, assuming no-slip boundary conditions at the surface of the cylinder, which is considered as large compared with the bilayer components (i.e., R Ͼ h). Numerous biological studies, both in model systems (2-4) and living cells (5, 6), refer to this continuum approach (7). Because D depends only weakly on R, the characterization of protein or rafts radii is delicate (8); for example, increasing the radius from 10 to 100 Å changes the mobility by a mere 30% [for h ϭ 30 Å and m ϭ 10 poise (P; 1 P ϭ 0.1 Pa⅐s)].To check the applicability of the Saffman-Delbrück formula (Eq. 1), we have used fringe pattern photobleaching under the microscope (9) to measure precisely the self-diffusion of transmembrane peptides and proteins of well characterized dimensions. Results and DiscussionThe weight of the bilayer thickness, h, has never been investigated. Rather than using lipids of various lengths, we opted for a unique system where the bilayer thickness can be continuously tuned, leaving the bilayer viscosity constant.We use a phase of model bilayers made of nonionic ...

show abstract

Size dependence of the translational diffusion of large integral membrane proteins in liquid-crystalline phase lipid bilayers. A study using fluorescence recovery after photobleaching

Cited by 126 publications

References 28 publications

Dopamine Transporter Activity Is Modulated by α-Synuclein

Dopamine Transporter Activity Is Modulated by α-Synuclein

A localized surface protein of guinea pig sperm exhibits free diffusion in its domain.

Lateral mobility of proteins in liquid membranes revisited

Contact Info

Product

Resources

About