Structural basis of the target-binding mode of the G protein-coupled receptor kinase-interacting protein in the regulation of focal adhesion dynamics

Liang, Mingfu1; Xie, Xingqiao1,2; Pan, Jian1; Jin, Gaowei1; Yu, Cong1,3,4; Wei, Zhiyi1,5

2019-04-12

10.1074/jbc.RA118.006915

JOURNAL OF BIOLOGICAL CHEMISTRY   影响因子和分区

1083351X

1083-351X

Focal adhesions (FAs) are specialized sites where intracellular cytoskeleton elements connect to the extracellular matrix and thereby control cell motility. FA assembly depends on various scaffold proteins, including the G protein-coupled receptor kinase-interacting protein 1 (GIT1), paxillin, and liprin-. Although liprin- and paxillin are known to competitively interact with GIT1, the molecular basis governing these interactions remains elusive. To uncover the underlying mechanisms of how GIT1 is involved in FA assembly by alternatively binding to liprin- and paxillin, here we solved the crystal structures of GIT1 in complex with liprin- and paxillin at 1.8 and 2.6 resolutions, respectively. These structures revealed that the paxillin-binding domain (PBD) of GIT1 employs distinct binding modes to recognize a single -helix of liprin- and the LD4 motif of paxillin. Structure-based design of protein variants produced two binding-deficient GIT1 variants; specifically, these variants lost the ability to interact with liprin- only or with both liprin- and paxillin. Expressing the GIT1 variants in COS7 cells, we discovered that the two PBD-meditated interactions play different roles in either recruiting GIT1 to FA or facilitating FA assembly. Additionally, we demonstrate that, unlike for the known binding mode of the FAT domain to LD motifs, the PBD of GIT1 uses different surface patches to achieve high selectivity in LD motif recognition. In summary, our results have uncovered the mechanisms by which GIT1's PBD recognizes cognate paxillin and liprin- structures, information we anticipate will be useful for future investigations of GIT1-protein interactions in cells.

focal adhesion crystal structure structure-function protein-protein interaction protein complex structural biology cell migration GTPase activating protein (GAP) complex structure paxillin (PXN) PTPRF-interacting protein alpha 2 (PPFIA2) single alpha-helix

SCI ; EI

英语

第一 ; 通讯

Shenzhen Science and Technology Innovation Commission[JCYJ20160229153100269] ; Shenzhen Science and Technology Innovation Commission[JCYJ20160301112450474] ; Shenzhen Science and Technology Innovation Commission[ZDSYS20140509142721429]

Biochemistry & Molecular Biology

Biochemistry & Molecular Biology

WOS:000465077500015

AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC

20191606805548

Adhesion ; Binding energy ; Budget control ; Enzymes

Biomedical Engineering:461.1 ; Physical Chemistry:801.4 ; Materials Science:951

BIOLOGY & BIOCHEMISTRY

Web of Science

1.Southern Univ Sci & Technol, Dept Biol, Shenzhen 518055, Peoples R China
2.Southern Univ Sci & Technol, Acad Adv Interdisciplinary Studies, Shenzhen 518055, Peoples R China
3.Guangdong Prov Key Lab Cell Microenvironm & Dis R, Shenzhen 518055, Peoples R China
4.Shenzhen Key Lab Cell Microenvironm, Shenzhen 518055, Peoples R China
5.SUSTech, Neural & Cognit Sci Res Ctr, Shenzhen, Peoples R China

Liang, Mingfu,Xie, Xingqiao,Pan, Jian,et al. Structural basis of the target-binding mode of the G protein-coupled receptor kinase-interacting protein in the regulation of focal adhesion dynamics[J]. JOURNAL OF BIOLOGICAL CHEMISTRY,2019,294(15):5827-5839.

Liang, Mingfu,Xie, Xingqiao,Pan, Jian,Jin, Gaowei,Yu, Cong,&Wei, Zhiyi.(2019).Structural basis of the target-binding mode of the G protein-coupled receptor kinase-interacting protein in the regulation of focal adhesion dynamics.JOURNAL OF BIOLOGICAL CHEMISTRY,294(15),5827-5839.

Liang, Mingfu,et al."Structural basis of the target-binding mode of the G protein-coupled receptor kinase-interacting protein in the regulation of focal adhesion dynamics".JOURNAL OF BIOLOGICAL CHEMISTRY 294.15(2019):5827-5839.