Structure and function of the intracellular region of the plexin-B1 transmembrane receptor

Yufeng Tong, Prasanta K. Hota, Junia Y. Penachioni, Mehdi B. Hamaneh, SoonJeung J. Kim, Rebecca S. Alviani, Limin Shen, Hao He, Wolfram Tempel, Luca Tamagnone, Hee Won Park, Matthias Buck

Research output: Contribution to journalArticle

54 Citations (Scopus)

Abstract

Members of the plexin family are unique transmembrane receptors in that they interact directly with Rho family small GTPases; moreover, they contain a GTPase-activating protein (GAP) domain for R-Ras, which is crucial for plexin-mediated regulation of cell motility. However, the functional role and structural basis of the interactions between the different intracellular domains of plexins remained unclear. Here we present the 2.4Å crystal structure of the complete intracellular region of human plexin-B1. The structure is monomeric and reveals that the GAP domain is folded into one structure from two segments, separated by the Rho GTPase binding domain (RBD). The RBD is not dimerized, as observed previously. Instead, binding of a conserved loop region appears to compete with dimerization and anchors the RBD to the GAP domain. Cell-based assays on mutant proteins confirm the functional importance of this coupling loop. Molecular modeling based on structural homology to p120GAP·H-Ras suggests that Ras GTPases can bind to the plexin GAP region. Experimentally, we show that the monomeric intracellular plexin-B1 binds R-Ras but not H-Ras. These findings suggest that the monomeric form of the intracellular region is primed for GAP activity and extend a model for plexin activation.

Original languageEnglish
Pages (from-to)35962-35972
Number of pages11
JournalJournal of Biological Chemistry
Volume284
Issue number51
DOIs
Publication statusPublished - Dec 18 2009

Fingerprint

GTPase-Activating Proteins
rho GTP-Binding Proteins
ras Proteins
Molecular modeling
Monomeric GTP-Binding Proteins
Dimerization
Mutant Proteins
Anchors
Cell Movement
plexin
Assays
Crystal structure
Chemical activation
Cells
Protein Domains

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Tong, Y., Hota, P. K., Penachioni, J. Y., Hamaneh, M. B., Kim, S. J., Alviani, R. S., ... Buck, M. (2009). Structure and function of the intracellular region of the plexin-B1 transmembrane receptor. Journal of Biological Chemistry, 284(51), 35962-35972. https://doi.org/10.1074/jbc.M109.056275

Structure and function of the intracellular region of the plexin-B1 transmembrane receptor. / Tong, Yufeng; Hota, Prasanta K.; Penachioni, Junia Y.; Hamaneh, Mehdi B.; Kim, SoonJeung J.; Alviani, Rebecca S.; Shen, Limin; He, Hao; Tempel, Wolfram; Tamagnone, Luca; Park, Hee Won; Buck, Matthias.

In: Journal of Biological Chemistry, Vol. 284, No. 51, 18.12.2009, p. 35962-35972.

Research output: Contribution to journalArticle

Tong, Y, Hota, PK, Penachioni, JY, Hamaneh, MB, Kim, SJ, Alviani, RS, Shen, L, He, H, Tempel, W, Tamagnone, L, Park, HW & Buck, M 2009, 'Structure and function of the intracellular region of the plexin-B1 transmembrane receptor', Journal of Biological Chemistry, vol. 284, no. 51, pp. 35962-35972. https://doi.org/10.1074/jbc.M109.056275
Tong Y, Hota PK, Penachioni JY, Hamaneh MB, Kim SJ, Alviani RS et al. Structure and function of the intracellular region of the plexin-B1 transmembrane receptor. Journal of Biological Chemistry. 2009 Dec 18;284(51):35962-35972. https://doi.org/10.1074/jbc.M109.056275
Tong, Yufeng ; Hota, Prasanta K. ; Penachioni, Junia Y. ; Hamaneh, Mehdi B. ; Kim, SoonJeung J. ; Alviani, Rebecca S. ; Shen, Limin ; He, Hao ; Tempel, Wolfram ; Tamagnone, Luca ; Park, Hee Won ; Buck, Matthias. / Structure and function of the intracellular region of the plexin-B1 transmembrane receptor. In: Journal of Biological Chemistry. 2009 ; Vol. 284, No. 51. pp. 35962-35972.
@article{80be9babd7dc4e99894e30ee9a1b8cc3,
title = "Structure and function of the intracellular region of the plexin-B1 transmembrane receptor",
abstract = "Members of the plexin family are unique transmembrane receptors in that they interact directly with Rho family small GTPases; moreover, they contain a GTPase-activating protein (GAP) domain for R-Ras, which is crucial for plexin-mediated regulation of cell motility. However, the functional role and structural basis of the interactions between the different intracellular domains of plexins remained unclear. Here we present the 2.4{\AA} crystal structure of the complete intracellular region of human plexin-B1. The structure is monomeric and reveals that the GAP domain is folded into one structure from two segments, separated by the Rho GTPase binding domain (RBD). The RBD is not dimerized, as observed previously. Instead, binding of a conserved loop region appears to compete with dimerization and anchors the RBD to the GAP domain. Cell-based assays on mutant proteins confirm the functional importance of this coupling loop. Molecular modeling based on structural homology to p120GAP·H-Ras suggests that Ras GTPases can bind to the plexin GAP region. Experimentally, we show that the monomeric intracellular plexin-B1 binds R-Ras but not H-Ras. These findings suggest that the monomeric form of the intracellular region is primed for GAP activity and extend a model for plexin activation.",
author = "Yufeng Tong and Hota, {Prasanta K.} and Penachioni, {Junia Y.} and Hamaneh, {Mehdi B.} and Kim, {SoonJeung J.} and Alviani, {Rebecca S.} and Limin Shen and Hao He and Wolfram Tempel and Luca Tamagnone and Park, {Hee Won} and Matthias Buck",
year = "2009",
month = "12",
day = "18",
doi = "10.1074/jbc.M109.056275",
language = "English",
volume = "284",
pages = "35962--35972",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "51",

}

TY - JOUR

T1 - Structure and function of the intracellular region of the plexin-B1 transmembrane receptor

AU - Tong, Yufeng

AU - Hota, Prasanta K.

AU - Penachioni, Junia Y.

AU - Hamaneh, Mehdi B.

AU - Kim, SoonJeung J.

AU - Alviani, Rebecca S.

AU - Shen, Limin

AU - He, Hao

AU - Tempel, Wolfram

AU - Tamagnone, Luca

AU - Park, Hee Won

AU - Buck, Matthias

PY - 2009/12/18

Y1 - 2009/12/18

N2 - Members of the plexin family are unique transmembrane receptors in that they interact directly with Rho family small GTPases; moreover, they contain a GTPase-activating protein (GAP) domain for R-Ras, which is crucial for plexin-mediated regulation of cell motility. However, the functional role and structural basis of the interactions between the different intracellular domains of plexins remained unclear. Here we present the 2.4Å crystal structure of the complete intracellular region of human plexin-B1. The structure is monomeric and reveals that the GAP domain is folded into one structure from two segments, separated by the Rho GTPase binding domain (RBD). The RBD is not dimerized, as observed previously. Instead, binding of a conserved loop region appears to compete with dimerization and anchors the RBD to the GAP domain. Cell-based assays on mutant proteins confirm the functional importance of this coupling loop. Molecular modeling based on structural homology to p120GAP·H-Ras suggests that Ras GTPases can bind to the plexin GAP region. Experimentally, we show that the monomeric intracellular plexin-B1 binds R-Ras but not H-Ras. These findings suggest that the monomeric form of the intracellular region is primed for GAP activity and extend a model for plexin activation.

AB - Members of the plexin family are unique transmembrane receptors in that they interact directly with Rho family small GTPases; moreover, they contain a GTPase-activating protein (GAP) domain for R-Ras, which is crucial for plexin-mediated regulation of cell motility. However, the functional role and structural basis of the interactions between the different intracellular domains of plexins remained unclear. Here we present the 2.4Å crystal structure of the complete intracellular region of human plexin-B1. The structure is monomeric and reveals that the GAP domain is folded into one structure from two segments, separated by the Rho GTPase binding domain (RBD). The RBD is not dimerized, as observed previously. Instead, binding of a conserved loop region appears to compete with dimerization and anchors the RBD to the GAP domain. Cell-based assays on mutant proteins confirm the functional importance of this coupling loop. Molecular modeling based on structural homology to p120GAP·H-Ras suggests that Ras GTPases can bind to the plexin GAP region. Experimentally, we show that the monomeric intracellular plexin-B1 binds R-Ras but not H-Ras. These findings suggest that the monomeric form of the intracellular region is primed for GAP activity and extend a model for plexin activation.

UR - http://www.scopus.com/inward/record.url?scp=72149087958&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=72149087958&partnerID=8YFLogxK

U2 - 10.1074/jbc.M109.056275

DO - 10.1074/jbc.M109.056275

M3 - Article

VL - 284

SP - 35962

EP - 35972

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 51

ER -