Genetic selection for and molecular dynamic modeling of a protein transmembrane domain multimerization motif from a random Escherichia coli genomic library

Research output: Contribution to journalArticlepeer-review

Authors

  • J A Leeds
  • D Boyd
  • G K Sonoda
  • H T Luu
  • D M Engelman
  • J Beckwith

Colleges, School and Institutes

Abstract

In order to identify new transmembrane helix packing motifs in naturally occurring proteins, we have selected transmembrane domains from a library of random Escherichia coli genomic DNA fragments and screened them for homomultimerization via their abilities to dimerize the bacteriophage lambda cI repressor DNA-binding domain. Sequences were isolated using a modified lambda cI headpiece dimerization assay system, which was shown previously to measure transmembrane helix-helix association in the E. coli inner membrane. Screening resulted in the identification of several novel sequences that appear to mediate helix-helix interactions. One sequence, representing the predicted sixth transmembrane domain (TM6) of the E. coli protein YjiO, was chosen for further analysis. Using site-directed mutagenesis and molecular dynamics, a small set of models for YjiO TM6 multimerization interface interactions were generated. This work demonstrates the utility of combining in vivo genetic tools with computational systems for understanding membrane protein structure and assembly.

Bibliographic note

Copyright 2001 Academic Press.

Details

Original languageEnglish
Pages (from-to)181-95
Number of pages15
JournalJournal of Molecular Biology
Volume313
Issue number1
Publication statusPublished - 12 Oct 2001

Keywords

  • Genes, Bacterial, Viral Proteins, Genomic Library, Protein Subunits, Amino Acid Motifs, Repressor Proteins, Genetic Vectors, Escherichia coli, Cell Membrane, Molecular Sequence Data, Amino Acid Substitution, Bacteriophage lambda, Viral Regulatory and Accessory Proteins, Models, Molecular, Dimerization, DNA-Binding Proteins, Amino Acid Sequence, Recombinant Fusion Proteins, Protein Sorting Signals, Membrane Proteins, Protein Binding, Cloning, Molecular, Binding Sites, Protein Structure, Quaternary, Base Sequence, Escherichia coli Proteins, Protein Structure, Tertiary, Protein Transport