西亚试剂：Tes, a Specific Mena Interacting Partner, Breaks the Rules

发布时间：2026-02-22

Tes, a Specific Mena Interacting Partner, Breaks the Rules for EVH1 Binding

Batiste Boëda,1,6 David C. Briggs,2,3,6 Theresa Higgins,1 Boyan K. Garvalov,4 Andrew J. Fadden,2 Neil Q. McDonald,2,5 and Michael Way1,

1 Cell Motility Laboratory, Cancer Research UK, London Research Institute, 44 Lincoln's Inn Fields, London, WC2A 3PX, UK
2 Structural Biology Laboratory, Cancer Research UK, London Research Institute, 44 Lincoln's Inn Fields, London, WC2A 3PX, UK
3 Wellcome Trust Centre for Cell-Matrix Research, University of Manchester, Manchester, M13 9PT, UK
4 Max Planck Institute of Neurobiology, Am Klopferspitz 18, D-82152 Martinsried, Germany
5 School of Crystallography, Birkbeck College, London, WC1E 7HX, UK

Corresponding author
Michael Way
michael.way@cancer.org.uk

Summary

The intracellular targeting of Ena/VASP family members is achieved via the interaction of their EVH1 domain with FPPPP sequence motifs found in a variety of cytoskeletal proteins, including lamellipodin, vinculin, and zyxin. Here we show that the LIM3 domain of Tes, which lacks the FPPPP motif, binds to the EVH1 domain of Mena, but not to those of VASP or Evl. The structure of the LIM3:EVH1 complex reveals that Tes occludes the FPPPP-binding site and competes with FPPPP-containing proteins for EVH1 binding. Structure-based gain-of-function experiments define the molecular basis for the specificity of the Tes-Mena interaction. Consistent with in vitro observations, the LIM3 domain displaces Mena, but not VASP, from the leading edge and focal adhesions. It also regulates cell migration through a Mena-dependent mechanism. Our observations identify Tes as an atypical EVH1 binding partner and a regulator specific to a single Ena/VASP family member.

 Figure S1. Tes Interacts Directly with the EVH1 Domain of Mena
(A) Immunoblot analysis of glutathione sepharose pull downs reveals that GST-EVH1 Mena resin retains GFP-tagged full length Tes (Tes) or its C-terminal half (C-Term, residues 230-421) but not the N-terminal half of the molecule (N-Term, residues 1-234) from HeLa cell extracts. The ponceau stain demonstrates all samples contained equivalent amounts of GST-EVH1 resin. The input (I) and bound (B) samples are indicated.
(B) Coomassie stained gel reveals that GST-EVH1 resin retains full length His-Tes but not His-Tes-C391A from a bacterial soluble fraction. The C391A mutation disrupts the structural integrity of the LIM3 domain and Mena binding (Garvalov et al, 2003).
(C) Immunoblot analysis of glutathione sepharose pull downs reveals that full length His-Tes is retained by a GST-EVH1 Mena resin but not by GST, GST-EVH2 Mena and GST-EVH1 VASP resins. The anti-GST immunoblot reveals that all samples contained similar amounts of each glutathione sepharose resin. In A-C, the protein on resin is
indicated with an asterisk.
(D) An example of the localization of Mena and vinculin in HeLa cells over expressing RFP-Tes or RFP-Tes-LIM3-4A.
(E) Quantification of Mena immunostaining at the focal adhesion of HeLa cells reveals that RFP-Tes but not RFP-Tes-LIM3-4A over expression significantly displaces Mena from focal adhesions. In each case 4x100 cells were analysed and the standard deviation is indicated. A standard student T-test gives a significance of p < 0.001 for the presence of Mena at focal adhesions between the two sets of data. Co-staining with anti-vinculin was used to assess the integrity of focal adhesions

• 以上资料由西亚试剂：http://www.xiyashiji.com/ 提供此产品的详细信息如密度,含量,分子式,分子量等均可在西亚官网查询   
• 相关产品如汞乙酸汞氯化汞氧化汞碘化汞硫酸汞硝酸汞溴化汞硝酸亚汞氯化亚汞乙酸苯汞碘化汞钾硫氰酸汞氯化氨基汞三氯生三氯氧磷三氯乙烯水合氯醛三氯化磷三氯化钌三氯化钛三氯化铱三氯化铑三氯硫磷三氯乙烷三氯甲烷三氯卡班TCC1,3,5-三氯苯1,2,4-三氯苯1,2,3-三氯苯无水氯化铝三氯乙酸酐三氯乙酸钠碘甲烷二碘甲烷三碘甲烷 三氟碘甲烷硫酸二甲酯氯磺酸苯硫酚苯硫酚钠3-氨基苯硫酚2,6-二氯苯硫酚2,4-二氯苯硫酚2,5-二氯苯硫酚2-甲氧基苯硫酚2-氯乙醇 等均有销售.欢迎订购

 

上一篇：西亚试剂：苯甲腈
下一篇：西亚试剂：Nanoscale architecture of integrin-based cell adhesions