Interaction of Mutant Alkaline Phosphatase Precursors with Membrane Phospholipids in vivo and in vitro

A. E. Kalinin, N. I. Mikhaleva, A. L. Karamyshev, Z. N. Karamysheva, M. A. Nesmeyanova

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Positively charged amino acid residues in the N-terminal domain of the signal peptides of secreted proteins are thought to interact with negatively charged anionic phospholipids during the initiation of secretion. To test this hypothesis, substitutions of the uncharged Ala or the negatively charged Glu residue for the positively charged Lys-20 of the N-terminus of the signal peptide of Escherichia coli alkaline phosphatase were introduced using a modified method of oligonucleotide-directed mutagenesis. We found that Lys-20 is involved in the interaction of the signal peptide with anionic phospholipids in vivo and effects the efficiency of insertion of the signal peptide of isolated precursor into model phospholipid membranes in vitro. We also show that the efficiency of signal peptide insertion into the lipid bilayer depends on the fluidity of the bilayer.

Original languageEnglish (US)
Pages (from-to)1021-1029
Number of pages9
JournalBiochemistry (Moscow)
Volume64
Issue number9
StatePublished - Sep 1999

Fingerprint

Protein Sorting Signals
Alkaline Phosphatase
Phospholipids
Membranes
Mutagenesis
Membrane Fluidity
Lipid bilayers
Fluidity
Lipid Bilayers
Site-Directed Mutagenesis
Oligonucleotides
Escherichia coli
Substitution reactions
In Vitro Techniques
Amino Acids
Proteins

Keywords

  • Alkaline phosphatase
  • Amino acid substitutions
  • E. Coli
  • Phospholipids
  • Protein translocation

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry

Cite this

Kalinin, A. E., Mikhaleva, N. I., Karamyshev, A. L., Karamysheva, Z. N., & Nesmeyanova, M. A. (1999). Interaction of Mutant Alkaline Phosphatase Precursors with Membrane Phospholipids in vivo and in vitro. Biochemistry (Moscow), 64(9), 1021-1029.

Interaction of Mutant Alkaline Phosphatase Precursors with Membrane Phospholipids in vivo and in vitro. / Kalinin, A. E.; Mikhaleva, N. I.; Karamyshev, A. L.; Karamysheva, Z. N.; Nesmeyanova, M. A.

In: Biochemistry (Moscow), Vol. 64, No. 9, 09.1999, p. 1021-1029.

Research output: Contribution to journalArticle

Kalinin, AE, Mikhaleva, NI, Karamyshev, AL, Karamysheva, ZN & Nesmeyanova, MA 1999, 'Interaction of Mutant Alkaline Phosphatase Precursors with Membrane Phospholipids in vivo and in vitro', Biochemistry (Moscow), vol. 64, no. 9, pp. 1021-1029.
Kalinin AE, Mikhaleva NI, Karamyshev AL, Karamysheva ZN, Nesmeyanova MA. Interaction of Mutant Alkaline Phosphatase Precursors with Membrane Phospholipids in vivo and in vitro. Biochemistry (Moscow). 1999 Sep;64(9):1021-1029.
Kalinin, A. E. ; Mikhaleva, N. I. ; Karamyshev, A. L. ; Karamysheva, Z. N. ; Nesmeyanova, M. A. / Interaction of Mutant Alkaline Phosphatase Precursors with Membrane Phospholipids in vivo and in vitro. In: Biochemistry (Moscow). 1999 ; Vol. 64, No. 9. pp. 1021-1029.
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