title: Probing for protein-protein interactions during cell migration: limitations and challenges.
abstract: Cellular migration is a fundamental biological process occurring as early as embryogenesis to the pathological state of cancer metastasis. Nearly all cellular migrations involve an extracellular signal that is transduced internally by members of a signalling cascade. These signal transduction events are driven by protein-protein interactions (PPIs) that coordinate intracellular activities to enable a cell to migrate. Understanding these PPIs will provide valuable insight into how cellular migration can be modulated perhaps towards a therapeutic end. Histologically, not many techniques are available to demonstrate PPIs. Contrasting agents only demonstrate the presence of a particular protein, and perhaps its co-localisation with another protein. Yet, co-localisation need not necessarily indicate physical interaction between the two proteins. In this review, we highlight four commonly used methods that continue to expand our understanding of PPIs underlying cell migration: co-immunoprecipitation, bimolecular fluorescence complementation, proximity ligation assay and surface plasmon resonance. The methods discussed herein allow for the study of PPIs in a wide variety of biological samples, including cell lysates, live cells, fixed cells and tissues, enabling the quantification of endogenous PPIs and exploration of the nature of PPIs. We also include a rudimentary framework for researchers to decide which experimental method best suits their research goals.
authors.affiliation: School of Biological Sciences, Nanyang Technological University, Singapore.
authors.forename: Jeremy Soon Kiat
authors.initials: JS
authors.lastname: Chan
authors.affiliation: School of Biological Sciences, Nanyang Technological University, Singapore.
authors.forename: Zi
authors.initials: Z
authors.lastname: Teo
authors.affiliation: School of Biological Sciences, Nanyang Technological University, Singapore.
authors.forename: Ming Keat
authors.initials: MK
authors.lastname: Sng
authors.affiliation: School of Biological Sciences, Nanyang Technological University, and Institute of Molecular and Cell Biology, Singapore. nstan@ntu.edu.sg or nstan@imcb.a-star.edu.sg.
authors.forename: Nguan Soon
authors.initials: NS
authors.lastname: Tan
mesh_terms: D000818:Animals; D002465:Cell Movement / Q000502:physiology*; D006801:Humans; D047468:Immunoprecipitation; D011485:Protein Binding / Q000502:physiology*; D011506:Proteins / Q000378:metabolism*
chemical_list: D011506:Proteins