Hydrophobic Matching Controls the Tilt and Stability of the Dimeric Platelet-derived Growth Factor Receptor (PDGFR) β Transmembrane Segment

Muhle‐Goll, Claudia; Hoffmann, Silke; Afonin, Sergii; Grage, Stephan L.; Polyansky, Anton A.; Windisch, Dirk; Zeitler, Marcel; Bürck, Jochen; Ulrich, Anne S.

doi:10.1074/jbc.m111.325555

Cited by 65 publications

(72 citation statements)

References 47 publications

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“…The interface between the two PDGFβR TMDs in the six-helix model is similar to the interface determined by NMR of a PDGFβR TMD dimer in the absence of the E5 protein (29). This NMR structure, in turn, appears consistent with the lowresolution structure of the full-length PDGFβR dimer activated by PDGF binding, as determined by negative-stain electron microscopy (15).…”

Section: Discussionsupporting

confidence: 70%

“…5A). If the PDGFβR TMDs in the active complex have a left-handed crossing angle [as suggested by the modeling conducted here and by NMR experiments on a PDGFβR peptide (29)], the substitutions in each mutant fall on a single face of the TMD. Mutations that lie in the PDGFβR/E5 interface are likely to disrupt the interaction of the E5 dimer(s) with the PDGFβR.…”

Section: Resultsmentioning

confidence: 99%

“…In addition to causing PDGFβR activation, complex formation between the E5 protein and the PDGFβR may help adapt the relatively long TMD of the PDGFβR to the thin lipid bilayer of the Golgi membrane (4,29).…”

Section: Significancementioning

confidence: 99%

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Two transmembrane dimers of the bovine papillomavirus E5 oncoprotein clamp the PDGF β receptor in an active dimeric conformation

Karabadzhak

Petti

Barrera

et al. 2017

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

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The dimeric 44-residue E5 protein of bovine papillomavirus is the smallest known naturally occurring oncoprotein. This transmembrane protein binds to the transmembrane domain (TMD) of the platelet-derived growth factor β receptor (PDGFβR), causing dimerization and activation of the receptor. Here, we use Rosetta membrane modeling and all-atom molecular dynamics simulations in a membrane environment to develop a chemically detailed model of the E5 protein/PDGFβR complex. In this model, an active dimer of the PDGFβR TMD is sandwiched between two dimers of the E5 protein. Biochemical experiments showed that the major PDGFβR TMD complex in mouse cells contains two E5 dimers and that binding the PDGFβR TMD to the E5 protein is necessary and sufficient to recruit both E5 dimers into the complex. These results demonstrate how E5 binding induces receptor dimerization and define a molecular mechanism of receptor activation based on specific interactions between TMDs.transmembrane protein complex | oncogene | traptamer | BPV | blue native gel electrophoresis B ecause viruses modulate signaling nodes that control cell behavior and virus replication, the study of viral proteins has provided great insight into many aspects of cellular biochemistry and the cellular processes that regulate biological function. Thus, viral proteins have long been recognized as valuable tools to probe central problems in biology. A particularly interesting viral protein is the 44-residue E5 oncoprotein encoded by bovine papillomavirus type 1 (BPV). The BPV E5 protein and closely related E5 proteins of other fibropapillomaviruses are the shortest known, naturally occurring proteins with tumorigenic potential (1). The E5 protein is an extremely hydrophobic integral membrane protein located primarily in the membranes of the Golgi apparatus of transformed cells (2, 3). Biophysical studies in model membranes indicate that the E5 protein adopts a transmembrane orientation roughly perpendicular to the membrane surface (4-6). In essence, the E5 protein is a freestanding transmembrane domain (TMD), with a type II transmembrane orientation in which a short C-terminal segment protrudes into the lumen of the Golgi (2). In cells, detergent micelles, and model lipid bilayers, the E5 protein exists as a homodimer stabilized by disulfide bonds involving C-terminal cysteine residues (3-5, 7-10). Genetic studies showed that E5 dimerization is required for transforming activity, and a preferred orientation of the E5 dimer with a symmetric homodimer interface has been identified (7,9,11,12).The E5 protein transforms cells by activating the cellular platelet-derived growth factor (PDGF) β receptor (PDGFβR), although there may be additional minor, alternative transforming pathways as well (13). The PDGFβR is a receptor tyrosine kinase (RTK) with an extracellular domain that binds PDGF, a hydrophobic membrane-spanning segment, and a cytoplasmic domain with tyrosine kinase activity. The inactive PDGFβR is primarily monomeric in unstimulated cells, and PDGF binding indu...

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Section: Discussionsupporting

confidence: 70%

Section: Resultsmentioning

confidence: 99%

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Two transmembrane dimers of the bovine papillomavirus E5 oncoprotein clamp the PDGF β receptor in an active dimeric conformation

Karabadzhak

Petti

Barrera

et al. 2017

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

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“…OCD is a powerful tool for studying the orientation of membrane-active peptides and protein segments in membranes (Lange et al, 2007;Bürck et al, 2008;Strandberg et al, 2008;Wadhwani et al, 2008Wadhwani et al, , 2012Wadhwani et al, , 2014Nolandt et al, 2009;Windisch et al, 2010;Heinzmann et al, 2011;Klein et al, 2012;Muhle-Goll et al, 2012;Steinbrecher et al, 2012;Aberle et al, 2014;Fanghänel et al, 2014). For α-helices, the OCD band at 208 nm can be used to estimate the helix tilt angle relative to the membrane normal (Wu et al, 1990).…”

Section: Orientation Of E5 In the Membrane By Ocdmentioning

confidence: 99%

Structural characterization of a C-terminally truncated E5 oncoprotein from papillomavirus in lipid bilayers

Windisch

Ziegler

Bürck

et al. 2014

Biological Chemistry

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E5 is the major transforming oncoprotein of bovine papillomavirus, which activates the platelet-derived growth factor receptor β in a highly specific manner. The short transmembrane protein E5 with only 44 residues interacts directly with the transmembrane segments of the receptor, but structural details are not available. Biophysical investigations are challenging, because the hydrophobic E5 protein tends to aggregate and get cross-linked non-specifically via two Cys residues near its C-terminus. Here, we demonstrate that a truncation by 10 amino acids creates a more manageable protein that can be conveniently used for structure analysis. Synchrotron radiation circular dichroism and solid-state (15)N- and (31)P-nuclear magnetic resonance spectroscopy show that this E5 variant serves as a representative model for the wild-type protein. The helical conformation of the transmembrane segment, its orientation in the lipid bilayer, and the ability to form homodimers in the membrane are not affected by the C-terminal truncation.

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“…The average M2 crossing angles are also decreased as the bilayer thickness increases, being 77.8 • in DLPC (n = 9), 66.5 • in POPC (n = 9) and 62.2 • in SOPC (n = 9). These data suggest that hydrophobic matching is the driving force behind changes in the tilt and crossing angles of M2 helices [1,20,2].…”

Section: The Packing Patterns Of L340 A344 and A347 In The Quaternarmentioning

confidence: 81%

Geometric rules of channel gating inferred from computational models of the P2X receptor transmembrane domain

2015

Journal of Molecular Graphics and Modelling

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Hydrophobic Matching Controls the Tilt and Stability of the Dimeric Platelet-derived Growth Factor Receptor (PDGFR) β Transmembrane Segment

Cited by 65 publications

References 47 publications

Two transmembrane dimers of the bovine papillomavirus E5 oncoprotein clamp the PDGF β receptor in an active dimeric conformation

Two transmembrane dimers of the bovine papillomavirus E5 oncoprotein clamp the PDGF β receptor in an active dimeric conformation

Structural characterization of a C-terminally truncated E5 oncoprotein from papillomavirus in lipid bilayers

Geometric rules of channel gating inferred from computational models of the P2X receptor transmembrane domain

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