title: A proteomic approach reveals integrin activation state-dependent control of microtubule cortical targeting.
abstract: Integrin activation, which is regulated by allosteric changes in receptor conformation, enables cellular responses to the chemical, mechanical and topological features of the extracellular microenvironment. A global view of how activation state converts the molecular composition of the region proximal to integrins into functional readouts is, however, lacking. Here, using conformation-specific monoclonal antibodies, we report the isolation of integrin activation state-dependent complexes and their characterization by mass spectrometry. Quantitative comparisons, integrating network, clustering, pathway and image analyses, define multiple functional protein modules enriched in a conformation-specific manner. Notably, active integrin complexes are specifically enriched for proteins associated with microtubule-based functions. Visualization of microtubules on micropatterned surfaces and live cell imaging demonstrate that active integrins establish an environment that stabilizes microtubules at the cell periphery. These data provide a resource for the interrogation of the global molecular connections that link integrin activation to adhesion signalling.
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, UK.
authors.forename: Adam
authors.initials: A
authors.lastname: Byron
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, UK.
authors.forename: Janet A
authors.initials: JA
authors.lastname: Askari
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, UK.
authors.forename: Jonathan D
authors.initials: JD
authors.lastname: Humphries
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, UK.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Jacquemet
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, UK.
authors.forename: Ewa J
authors.initials: EJ
authors.lastname: Koper
authors.affiliation: Biological Mass Spectrometry Core Facility, Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, UK.
authors.forename: Stacey
authors.initials: S
authors.lastname: Warwood
authors.affiliation: 1] Department of Biomedical Engineering, Boston University, Boston, Massachusetts 02215, USA [2] Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts 02115, USA.
authors.forename: Colin K
authors.initials: CK
authors.lastname: Choi
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, UK.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Stroud
authors.affiliation: 1] Department of Biomedical Engineering, Boston University, Boston, Massachusetts 02215, USA [2] Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts 02115, USA.
authors.forename: Christopher S
authors.initials: CS
authors.lastname: Chen
authors.affiliation: Biological Mass Spectrometry Core Facility, Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, UK.
authors.forename: David
authors.initials: D
authors.lastname: Knight
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, UK.
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Humphries
mesh_terms: D000495:Allosteric Site; D000911:Antibodies, Monoclonal / Q000737:chemistry; D002540:Cerebral Cortex / Q000378:metabolism; D016000:Cluster Analysis; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D005347:Fibroblasts / Q000378:metabolism; D052816:Foreskin / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D019012:Integrin beta1 / Q000378:metabolism; D016023:Integrins / Q000378:metabolism*; D020014:K562 Cells; D008297:Male; D013058:Mass Spectrometry; D008856:Microscopy, Fluorescence; D008870:Microtubules / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D015398:Signal Transduction
chemical_list: D000911:Antibodies, Monoclonal; D004129:Dimethylpolysiloxanes; D019012:Integrin beta1; D016023:Integrins; D020543:Proteome; D049452:Green Fluorescent Proteins; C013830:baysilon