β-Galactosidase fused to the hydrophobic domain of cytochrome b5 spontaneously associates with liposomes

Sathish K. George, Lydia Najera, Ramona Sandoval, Garret M. Ihler

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

Since liver microsomal cytochrome b5 spontaneously associates with liposomes and membranes by means of its C-terminal hydrophobic domain (HP), chimeric proteins containing HP prepared by genetic fusion might also spontaneously associate with liposomes or cellular membranes. Synthetic DNA corresponding to the hydrophobic domain of cytochrome b5 was enzymatically fused in-frame to cloned DNA corresponding to the C-terminus of the Escherichia coli enzyme, β-galactosidase. This protein, LacZ:HP, synthesized in E. coli and purified from a crude E. coli membrane extract, was shown to spontaneously associate with liposomes, as does cytochrome b5. Association is rapid and stable in the presence of salt and high pH and the fusion protein behaves as an integral membrane protein. LacZ:HP can be readily and extensively purified from crude extracts by association with liposomes and this procedure may provide a convenient purification scheme for proteins not otherwise readily purified, for example polypeptides from cloned gene fragments to be used for antibody production. These hybrid proteins may represent a new potentially useful class of polypeptides capable of hydrophobic interactions with membranes.

Original languageEnglish (US)
Pages (from-to)26-32
Number of pages7
JournalBBA - Biomembranes
Volume1061
Issue number1
DOIs
StatePublished - Jan 9 1991

Bibliographical note

Funding Information:
This work was supported in part by a grant from The Texas Advanced Technology Program (Universal Membrane Insertion Polypeptide, 1/86-8/89) and by funds from the Tom and Jean McMullin Chair of Genetics.

Keywords

  • Chimeric protein
  • Cytochrome b
  • Fusion protein
  • Hydrophobic domain
  • Liposome
  • Phosphatidylcholine
  • β-Galactosidase

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