Secondary Structure Analysis of a Functional Construct of Caveolin-1 Reveals a Long C-Terminal Helix

Plucinsky, Sarah M.; Glover, Kerney Jebrell

doi:10.1016/j.bpj.2015.08.030

Cited by 23 publications

(31 citation statements)

References 22 publications

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“…Our results also demonstrate the presence of helical structure within the C-terminal domain, consistent with what has previously been observed for Cav1 (27). However, it should be noted that we observed four different helical segments within the C-terminal domain of Cav3 (helix 3, residues 106-113, helix 4, residues 117-120, helix 5, residues 125-128, and helix 6, residues 132-146) as opposed to one long C-terminal helix observed in nonlipidated Cav1 (27). The CSI values proximal to the lipidation sites in Cav3 (residues 114-116 and 129-131) are low compared to those values reported for the corresponding residues in Cav1 (Fig.…”

Section: Discussionsupporting

confidence: 92%

“…Our analysis of the full-length lipidated form of Cav3 indicates that the scaffolding domain consists of a long membrane-associated a-helix that is contiguous with the helix-break-helix motif within the caveolin membrane domain (25,26). Our results also demonstrate the presence of helical structure within the C-terminal domain, consistent with what has previously been observed for Cav1 (27). However, it should be noted that we observed four different helical segments within the C-terminal domain of Cav3 (helix 3, residues 106-113, helix 4, residues 117-120, helix 5, residues 125-128, and helix 6, residues 132-146) as opposed to one long C-terminal helix observed in nonlipidated Cav1 (27).…”

Section: Discussionsupporting

confidence: 91%

“…The CSI values proximal to the lipidation sites in Cav3 (residues 114-116 and 129-131) are low compared to those values reported for the corresponding residues in Cav1 (Fig. S2) (27). Furthermore, a comparison of the CSI values of lipidated and nonlipidated Cav3 reveals differences proximal to the lipidation sites (Fig.…”

Section: Discussionmentioning

confidence: 86%

“…The membrane-embedded domain consists of a helix-break-helix motif that enters and exits the inner membrane leaflet (25,26). Finally, the cytosolic C-terminal domain of Cav1 was recently found to contain one long a-helix (27). This domain also contains three cysteine residues (residues 106, 116, and 129) known to undergo palmitoylation in Cav1 (28,29).…”

Section: Introductionmentioning

confidence: 98%

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A pH-Mediated Topological Switch within the N-Terminal Domain of Human Caveolin-3

Kim

Schlebach

et al. 2016

Biophysical Journal

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Caveolins mediate the formation of caveolae, which are small omega-shaped membrane invaginations involved in a variety of cellular processes. There are three caveolin isoforms, the third of which (Cav3) is expressed in smooth and skeletal muscles. Mutations in Cav3 cause a variety of human muscular diseases. In this work, we characterize the secondary structure, dynamics, and topology of the monomeric form of the full-length lipidated protein. Cav3 consists of a series of membrane-embedded or surface-associated helical elements connected by extramembrane connecting loops or disordered domains. Our results also reveal that the N-terminal domain undergoes a large scale pH-mediated topological rearrangement between soluble and membrane-anchored forms. Considering that roughly one-third of pathogenic mutations in Cav3 influence charged residues located in this domain, we hypothesize that this transition is likely to be relevant to the molecular basis of Cav3-linked diseases. These results provide insight into the structure of Cav3 and set the stage for mechanistic investigations of the effects of pathogenic mutations.

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Section: Discussionsupporting

confidence: 92%

Section: Discussionsupporting

confidence: 91%

Section: Discussionmentioning

confidence: 86%

Section: Introductionmentioning

confidence: 98%

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A pH-Mediated Topological Switch within the N-Terminal Domain of Human Caveolin-3

Kim

Schlebach

et al. 2016

Biophysical Journal

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“…Studies by many laboratories in recent years are now beginning to bridge this gulf using approaches that include biochemical isolation, NMR spectroscopy, EM, and X-ray crystallography (6,10,(12)(13)(14)(15)(16)(17). A picture is now emerging of a vesicular structure that possesses much greater complexity than originally thought.…”

mentioning

confidence: 99%

Unraveling the architecture of caveolae

Parton

Collins

2016

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

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The eukaryotic cell surface is composed of many distinct membrane domains that are formed by the cooperative interactions of different proteins and lipids. These domains are important for membrane trafficking and cell signaling and are modulated in turn by changes in the cell environment. Caveolae ("little caves") are ∼60-nm membrane invaginations ( Fig. 1) that are a dominant surface feature of many mammalian cells, including muscle fibers, endothelia, and adipocytes, where they play a role in membrane homeostasis, signaling, and cellular mechanoprotection. Formation of caveolae in vertebrate cells requires two distinct protein families: the membrane-embedded caveolins (CAV1-3) and the peripheral membrane cavins (Cavin1-4). Although the general morphology of caveolae has been known for decades, the atypical structures of the protein subunits has meant that progress has been slow with regards to the high-resolution studies of caveola architecture. By high-resolution scanning electron microscopy (EM) and frozen deepetch transmission EM, caveolae have been shown to be coated with striations (1, 2) or to possess spike-like structures (3), very different from other well-characterized vesicle coats, such as clathrin (4, 5). In PNAS, Stoeber et al. use a combination of biochemical dissection and EM to provide important insights into the underlying architecture of the caveola protein coat (6).The first protein component of caveolae to be identified and shown to be essential for caveola formation was CAV1 (7-9). Subsequently, it was found that both CAV2 and muscle-specific CAV3 homologs are also localized to caveolae. Caveolin proteins are ∼20 kDa and possess two membrane-inserted α-helices with cytoplasmic N-and C-terminal sequences. These proteins form oligomeric complexes at the plasma membrane that line the cytoplasmic leaflet of the invaginated caveola structures. Cavins, although equally important for caveola formation, were only identified in the past decade through proteomic and cellular studies from several groups. Cavins also possess a conserved and unique structural organization, in this case consisting of conserved N-and C-terminal α-helical domains (HR1 and HR2) interspersed by disordered linker sequences (DR1, DR2, DR3) (10). Although recent studies have begun to tease out the functions of these domains in membrane recruitment, remodeling, and protein-protein interactions, our understanding is still limited. For example, how do caveolin oligomers assemble within caveolae, and what drives cavin oligomerization and recruitment to caveolar domains at the cell surface?In their report, Stoeber et al. (6) propose a model for the arrangement of the caveolin and cavin proteins that begins to address some of the ongoing questions in the field. Affinity purification of Cavin1 expressed in HEK293 cells revealed the presence of heterogeneous particles with a mesh-like appearance by cryoEM and 3D electron tomography. These particles bound robustly to liposomes containing phosphatidylserine (PS) or phosphatidic acid, as...

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Total Synthesis and Structural Characterization of Caveolin‐1

et al. 2021

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Caveolin-1, which is an essential protein for caveola formation, was chemically synthesized. It is composed of 177 amino acid residues, is triply palmitoylated at the C-terminal region, and is inserted into the lipid bilayer to form a V-shaped structure in the middle of the polypeptide chain. The entire sequence was divided into five peptide segments, each of which was synthesized by the solid-phase method. To improve the solubility of the C-terminal region, O-acyl isopeptide structures were incorporated. After ligation by the thioester method and the introduction of the palmitoyl groups, all the protecting groups were removed and the isopeptide structures were converted into the native peptide bond. Finally, the obtained polypeptide was successfully inserted into bicelles, thus showing the success of the synthesis.

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Secondary Structure Analysis of a Functional Construct of Caveolin-1 Reveals a Long C-Terminal Helix

Cited by 23 publications

References 22 publications

A pH-Mediated Topological Switch within the N-Terminal Domain of Human Caveolin-3

A pH-Mediated Topological Switch within the N-Terminal Domain of Human Caveolin-3

Unraveling the architecture of caveolae

Total Synthesis and Structural Characterization of Caveolin‐1

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