A method for probing the topography and interactions of proteins: Footprinting of myoglobin

Min Zhong, Laura Lin, Neville R. Kallenbach

Research output: Contribution to journalArticlepeer-review

Abstract

We describe a procedure for mapping residues on the surface of a protein molecule to its sequence, using a scheme that is analogous to nucleic acid footprinting. The protein is end labeled radioactively and subjected to limited proteolysis, and the products are analyzed by denaturing polyacrylamide gel electrophoresis and autoradiography. The method is tested with the heme protein myoglobin and applied to mapping the (unknown) surface of the molecule lacking the heme group: apomyoglobin. Sites of protein- protein interaction can be identified, as illustrated by footprinting the association between myoglobin and an anti-myoglobin monoclonal antibody.

Original languageEnglish (US)
Pages (from-to)2111-2115
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume92
Issue number6
DOIs
StatePublished - Mar 14 1995

Keywords

  • protein-protein interaction
  • surface mapping

ASJC Scopus subject areas

  • General

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