Excursion of a single polypeptide into a protein pore: Simple physics, but complicated biology

Mohammad M. Mohammad, Liviu Movileanu

Research output: Contribution to journalArticle

57 Scopus citations

Abstract

Despite its fundamental and critical importance in molecular biology and practical medical biotechnology, how a polypeptide interacts with a transmembrane protein pore is not yet comprehensively understood. Here, we employed single-channel electrical recordings to reveal the interactions of short polypeptides and small folded proteins with a robust β-barrel protein pore. The short polypeptides were ∼25 residues in length, resembling positively charged targeting presequences involved in protein import. The proteins were consisted of positively charged pre-cytochrome b 2 fragments (pb2) fused to the small ribonuclease barnase (∼110 residues, Ba). Single-molecule experiments exploring the interaction of a folded pb2-Ba protein with a single β-barrel pore, which contained negatively charged electrostatic traps, revealed the complexity of a network of intermolecular forces, including driving and electrostatic ones. In addition, the interaction was dependent on other factors, such as the hydrophobic content of the interacting polypeptide, the location of the electrostatic trap, the length of the pb2 presequence and temperature. This single-molecule approach together with protein design of either the interacting polypeptide or the pore lumen opens new opportunities for the exploration of the polypeptide-pore interaction at high temporal resolution. Such future studies are also expected to unravel the advantages and limitations of the nanopore technique for the detection and exploration of individual polypeptides.

Original languageEnglish (US)
Pages (from-to)913-925
Number of pages13
JournalEuropean Biophysics Journal
Volume37
Issue number6
DOIs
StatePublished - Jul 1 2008

Keywords

  • Electrophysiology
  • Electrostatic interaction
  • Presequence
  • Protein design
  • Protein translocation
  • Single-molecule biophysics

ASJC Scopus subject areas

  • Biophysics

Fingerprint Dive into the research topics of 'Excursion of a single polypeptide into a protein pore: Simple physics, but complicated biology'. Together they form a unique fingerprint.

  • Cite this