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Engineering Synthetic Antibody Inhibitors Specific for LD2 or LD4 Motifs of Paxillin.

Nocula-Lugowska M, Lugowski M, Salgia R, Kossiakoff AA
J Mol Biol 427 2532-2547 (2015)
Cited: 8 times
EuropePMC logo PMID: 26087144

Abstract

Focal adhesion protein paxillin links integrin and growth factor signaling to actin cytoskeleton. Most of paxillin signaling activity is regulated via leucine-rich LD motifs (LD1-LD5) located at the N-terminus. Here, we demonstrate a method to engineer highly selective synthetic antibodies (sABs) against LD2 and LD4 that are binding sites for focal adhesion kinase (FAK) and other proteins. Phage display selections against peptides were used to generate sABs recognizing each LD motif. In the obtained X-ray crystal structures of the LD-sAB complexes, the LD motifs are helical and bind sABs through a hydrophobic side, similarly as in the structures with natural paxillin partners. The sABs are capable of pulling down endogenous paxillin in complex with FAK and can visualize paxillin in focal adhesions in cells. They were also used as selective inhibitors to effectively compete with focal adhesion targeting domain of FAK for the binding to LD2 and LD4. The sABs are tools for investigation of paxillin LD binding "platforms" and are capable of inhibiting paxillin interactions, thereby useful as potential therapeutics in the future.

Articles - 4xgz mentioned but not cited (3)

  1. Mechanism of integrin activation by talin and its cooperation with kindlin. Lu F, Zhu L, Bromberger T, Yang J, Yang Q, Liu J, Plow EF, Moser M, Qin J. Nat Commun 13 2362 (2022)
  2. Engineering Synthetic Antibody Inhibitors Specific for LD2 or LD4 Motifs of Paxillin. Nocula-Lugowska M, Lugowski M, Salgia R, Kossiakoff AA. J Mol Biol 427 2532-2547 (2015)
  3. Quaternary structure independent folding of voltage-gated ion channel pore domain subunits. Arrigoni C, Lolicato M, Shaya D, Rohaim A, Findeisen F, Fong LK, Colleran CM, Dominik P, Kim SS, Schuermann JP, DeGrado WF, Grabe M, Kossiakoff AA, Minor DL. Nat Struct Mol Biol 29 537-548 (2022)


Reviews citing this publication (3)

  1. The "Sticky Patch" Model of Crystallization and Modification of Proteins for Enhanced Crystallizability. Derewenda ZS, Godzik A. Methods Mol Biol 1607 77-115 (2017)
  2. Protein Engineering: Advances in Phage Display for Basic Science and Medical Research. Davydova EK. Biochemistry (Mosc) 87 S146-S110 (2022)
  3. The Role of Paxillin Aberrant Expression in Cancer and Its Potential as a Target for Cancer Therapy. Liu W, Huang X, Luo W, Liu X, Chen W. Int J Mol Sci 24 8245 (2023)

Articles citing this publication (2)

  1. Paxillin family of focal adhesion adaptor proteins and regulation of cancer cell invasion. Alpha KM, Xu W, Turner CE. Int Rev Cell Mol Biol 355 1-52 (2020)
  2. Synthetic Antibodies Detect Distinct Cellular States of Chromosome Passenger Complex Proteins. Ura M, Mukherjee S, Marcon E, Koestler SA, Kossiakoff AA. J Mol Biol 434 167602 (2022)


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