Preferred SH3 Domain Partners of ADAM Metalloproteases Include Shared and ADAM-Specific SH3 Interactions

Iivari Kleino,Jussi Hepojoki,Kalle Saksela,Ari Pekka Huovila,Annika Järviluoma,Laszlo Buday

doi:10.1371/journal.pone.0121301

Iivari Kleino, Jussi Hepojoki + Show 4 more

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https://doi.org/10.1371/journal.pone.0121301

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Abstract

A disintegrin and metalloproteinases (ADAMs) constitute a protein family essential for extracellular signaling and regulation of cell adhesion. Catalytic activity of ADAMs and their predicted potential for Src-homology 3 (SH3) domain binding show a strong correlation. Here we present a comprehensive characterization of SH3 binding capacity and preferences of the catalytically active ADAMs 8, 9, 10, 12, 15, 17, and 19. Our results revealed several novel interactions, and also confirmed many previously reported ones. Many of the identified SH3 interaction partners were shared by several ADAMs, whereas some were ADAM-specific. Most of the ADAM-interacting SH3 proteins were adapter proteins or kinases, typically associated with sorting and endocytosis. Novel SH3 interactions revealed in this study include TOCA1 and CIP4 as preferred partners of ADAM8, and RIMBP1 as a partner of ADAM19. Our results suggest that common as well as distinct mechanisms are involved in regulation and execution of ADAM signaling, and provide a useful framework for addressing the pathways that connect ADAMs to normal and aberrant cell behavior.

Highlights

A disintegrin and metalloproteinases (ADAMs)-mediated protein ectodomain cleavage—dubbed shedding—provides means to rapidly modify the amount and function of proteins on the cell surface
Presence of putative Src-homology 3 (SH3) domain binding motifs is a prominent feature of the cytosolic tails of ADAMs with catalytic activity
The peptide data could not be used to predict the SH3 interactions of the complete ADAM tails revealed by the phage library screening

Summary

Introduction

ADAM-mediated protein ectodomain cleavage—dubbed shedding—provides means to rapidly modify the amount and function of proteins on the cell surface. ADAM substrates range from growth factors and cytokines to receptors and adhesion proteins Their cleavage regulates or initiates many cellular processes including cytokine and growth factor signaling, cell adhesion, cell migration, and release of intracellular signaling domains from membrane bound precursor proteins. ADAM17 with over a hundred identified substrates, and ADAM10 with seventy, are the primary sheddases responsible for most of the ADAM-mediated ectodomain shedding characterized so far [1]. They share more than half of the substrates, usually shedding by ADAM10 or ADAM17 is initiated by different stimuli [3,4]. In many cases ADAM10 is either constantly shedding or activated by Ca2+ influx, whereas ADAM17 can be activated with

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