Single-molecule Force Spectroscopy of Protein-membrane Interactions

Overview

Journal Elife

Specialty Biology

Date 2017 Oct 31

PMID 29083305

Citations 27

Authors

Lu Ma

Yiying Cai

Yanghui Li

Junyi Jiao

Zhenyong Wu

Ben OShaughnessy

Pietro De Camilli

Erdem Karatekin

Yongli Zhang

Affiliations

Soon will be listed here.

Abstract

Many biological processes rely on protein-membrane interactions in the presence of mechanical forces, yet high resolution methods to quantify such interactions are lacking. Here, we describe a single-molecule force spectroscopy approach to quantify membrane binding of C2 domains in Synaptotagmin-1 (Syt1) and Extended Synaptotagmin-2 (E-Syt2). Syts and E-Syts bind the plasma membrane via multiple C2 domains, bridging the plasma membrane with synaptic vesicles or endoplasmic reticulum to regulate membrane fusion or lipid exchange, respectively. In our approach, single proteins attached to membranes supported on silica beads are pulled by optical tweezers, allowing membrane binding and unbinding transitions to be measured with unprecedented spatiotemporal resolution. C2 domains from either protein resisted unbinding forces of 2-7 pN and had binding energies of 4-14 kT per C2 domain. Regulation by bilayer composition or Ca recapitulated known properties of both proteins. The method can be widely applied to study protein-membrane interactions.

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