Caveolae are highly-curved invaginated micro-domains located in the plasma membrane that play a central role in a variety of cellular processes. Caveolins (1, 2, and 3) are the most important proteins found in caveolae, and are responsible for giving caveolae their unusual flask-like shape. Recent evidence has shown that improper regulation and mutant forms of caveolin can result in a variety of diseases including Alzheimer's, muscular dystrophy, cancer, and heart disease. Caveolin adopts an unusual intra-membrane horseshoe conformation where both its N- and C-termini face the cytoplasm, and this conformation is thought to promote membrane curvature. In addition, via high-order oligomerization, caveolin forms a structural backbone which stabilizes the membrane curvature. Using biophysical techniques such as nuclear magnetic resonance (NMR), fluorescence spectroscopy, and analytical ultracentrifugation, our objective is to characterize caveolin-1 on a fundamental level. This will be achieved by pursuing the following two specific aims: 1. Investigation of the membrane topology and three-dimensional structure of caveolin-1. 2. Investigation of caveolin-1 oligomerization.
Specific aim 1 will determine the high-resolution three-dimensional solution structure of caveolin-1 as well as examine the solvent accessibility of tryptophan residues to assess the topology of caveolin-1 in a bilayer. Next, the role that two conserved proline residues play in the creation and/or stabilization of the intra-membrane horseshoe conformation will be probed using site-directed mutagenesis.
Specific aim 2 will characterize both the size and distribution of oligomers formed by caveolin-1 in the presence and absence of cholesterol. Additionally, the role that a proline to leucine mutant plays in the oligomerization process will be probed. A fundamental understanding of caveolin-1 structure and oligomerization will undoubtedly open the door to possible therapeutic interventions that could address diseases linked to caveolin misfunction.
Caveolins are proteins that play an important role in a variety of events that happen within the cell. Alterations in the behavior of caveolins have been implicated in a variety of diseases including Alzheimer's, muscular dystrophy, cancer, and heart disease. Therefore, a more complete understanding of caveolins is necessary.
|Plucinsky, Sarah M; Root, Kyle T; Glover, Kerney Jebrell (2018) Efficient solubilization and purification of highly insoluble membrane proteins expressed as inclusion bodies using perfluorooctanoic acid. Protein Expr Purif 143:34-37|
|Mukai, Masaru; Glover, Kerney Jebrell; Regen, Steven L (2016) Evidence for Surface Recognition by a Cholesterol-Recognition Peptide. Biophys J 110:2577-80|
|Root, Kyle T; Glover, Kerney Jebrell (2016) Reconstitution and spectroscopic analysis of caveolin-1 residues 62-178 reveals that proline 110 governs its structure and solvent exposure. Biochim Biophys Acta 1858:682-8|
|Plucinsky, Sarah M; Glover, Kerney Jebrell (2015) Secondary Structure Analysis of a Functional Construct of Caveolin-1 Reveals a Long C-Terminal Helix. Biophys J 109:1686-8|
|Rui, Huan; Root, Kyle T; Lee, Jinwoo et al. (2014) Probing the U-shaped conformation of caveolin-1 in a bilayer. Biophys J 106:1371-80|
|Rieth, Monica D; Lee, Jinwoo; Glover, Kerney Jebrell (2012) Probing the caveolin-1 P132L mutant: critical insights into its oligomeric behavior and structure. Biochemistry 51:3911-8|
|Lee, Jinwoo; Glover, Kerney Jebrell (2012) The transmembrane domain of caveolin-1 exhibits a helix-break-helix structure. Biochim Biophys Acta 1818:1158-64|