Journal
JOURNAL OF PHYSICAL CHEMISTRY B
Volume 116, Issue 17, Pages 5122-5131Publisher
AMER CHEMICAL SOC
DOI: 10.1021/jp210045r
Keywords
-
Categories
Funding
- Office of Science, Office of Basic Energy Sciences, Chemical Sciences, Geosciences, and Biosciences Division of the U.S. Department of Energy (DOE) [DE-AC02-05CH11231]
- EPSCoR
- Office Of The Director [814251] Funding Source: National Science Foundation
Ask authors/readers for more resources
Transient recruitment of proteins to membranes is a fundamental mechanism by which the cell exerts spatial and temporal control over proteins' localization and interactions. Thus, the specificity and the kinetics of peripheral proteins' membrane residence are an attribute of their function. Here, we describe the membrane interactions of the interfacial epsin N-terminal homology (ENTH) domain with its target lipid phosphatidylinositol (4,5)-bisphosphate (PtdIns(4,5)P-2). The direct visualization and quantification of interactions of single ENTH molecules with supported lipid bilayers is achieved using total internal reflection fluorescence microscopy (TIRFM) with a time resolution of 13 ms. This enables the recording of the kinetic behavior of ENTH interacting with membranes with physiologically relevant concentrations of PtdIns(4,5)P-2 despite the low effective binding affinity. Subsequent single fluorophore tracking permits us to build up distributions of residence times and to measure ENTH dissociation rates as a function of membrane composition. Furthermore, due to the high time resolution, we are able to resolve details of the motion of ENTH associated with a simple, homogeneous membrane. In this case ENTH's diffusive transport appears to be the result of at least three different diffusion processes.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available