Membrane transport proteins with complete replacement of transmembrane helices with polyalanine sequences remain functional

Xiaoyong Bao, Yongyue Chen, Haeng Lee Sung, Chang Lee Sung, Luis Reuss, Guillermo A. Altenberg

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Approximately 25% of all genome coding sequences correspond to membrane proteins, which perform varied and essential functions in cells. Eukaryotic integral membrane proteins are predominantly α-helical proteins that span the membrane several times. The most frequent approach to identifying transmembrane-helix amino acids essential for function is to substitute native residues, one at a time, with Cys or Ala (Cys- and Alascanning mutagenesis). Here, we present a new approach, in which complete transmembrane-helix native sequences are substituted with poly-Ala sequences. We show that the basic functional features of two dissimilar membrane proteins, which function as a channel and a pump, respectively, are maintained when certain individual α-helices are replaced with poly-Ala sequences. This approach ("helix-scanning mutagenesis") allows for rapid identification of helices containing residues essential for function and can be used as a primary helix-screening tool, followed by individual amino acid substitutions when specific helix poly-Ala replacements cause functional changes in the protein.

Original languageEnglish (US)
Pages (from-to)8647-8650
Number of pages4
JournalJournal of Biological Chemistry
Volume280
Issue number10
DOIs
StatePublished - Mar 11 2005

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Membrane Transport Proteins
Mutagenesis
Membrane Proteins
Essential Amino Acids
Screening
Proteins
Substitution reactions
Amino Acid Substitution
Genes
Pumps
Membranes
Scanning
Amino Acids
Genome
polyalanine

ASJC Scopus subject areas

  • Biochemistry

Cite this

Membrane transport proteins with complete replacement of transmembrane helices with polyalanine sequences remain functional. / Bao, Xiaoyong; Chen, Yongyue; Sung, Haeng Lee; Sung, Chang Lee; Reuss, Luis; Altenberg, Guillermo A.

In: Journal of Biological Chemistry, Vol. 280, No. 10, 11.03.2005, p. 8647-8650.

Research output: Contribution to journalArticle

Bao, Xiaoyong ; Chen, Yongyue ; Sung, Haeng Lee ; Sung, Chang Lee ; Reuss, Luis ; Altenberg, Guillermo A. / Membrane transport proteins with complete replacement of transmembrane helices with polyalanine sequences remain functional. In: Journal of Biological Chemistry. 2005 ; Vol. 280, No. 10. pp. 8647-8650.
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