Protocol

Protein Interactions Captured by Chemical Cross-linking: Simple Cross-linking Screen Using Sulfo-MBS

This protocol was adapted from "Protein Interactions Captured by Chemical Cross-linking," Chapter 7, in Protein-Protein Interactions (eds. Golemis and Adams). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2005.

INTRODUCTION

This protocol describes a method for chemical cross-linking of proteins using sulfo-MBS (m-maleimidobenzoyl-N-hydroxysulfo-succinimide ester). Optimal conditions for cross-linking can be determined rapidly for a fixed concentration of a protein complex by varying the time of cross-linking, pH of the reaction, and concentration of sulfo-MBS. Typically, these screens require only small amounts of target proteins and can be carried out in less than a day.