The study of biogenesis and secretion of alkaline phoshatase and its mutant forms in Escherichia coli. III. Substitutions of N-terminal amino acid of alkaline phosphatase affect its biogenesis

A. L. Karamyshev, A. E. Kalinin, M. I. Khmelnitsky, M. G. Shlyapnikov, V. N. Ksenzenko, M. A. Nesmeyanova

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4 Scopus citations

Abstract

The effect of the N-terminal amino acid substitution on E. coli alkaline phosphatase biogenesis has been studied. The substitutions of Ser, Gln, Tyr, Leu, Gly, Ala, Glu, Phe, His, Cys, Lys and Pro for Arg(+1) were obtained by creating amber mutation at the corresponding position within phoA gene and expressing this mutated gene in E. coli strains that produce the amber-suppressor tRNAs. All mutant proteins were shown to translocate across the cytoplasmic membrane and possess enzyme activity. The introduction of Pro in +1 position disturbs the cleavage of signal peptide whereas the insertion of the other amino acids does not change the rates of processing in comparison with wild-type protein. All amino acid substitutions affect alkaline phosphatase isoenzyme composition. Some experimental evidence were also obtained on the specificity of protease, which split off N-terminal Arg during alkaline phosphatase maturation.

Original languageEnglish (US)
Pages (from-to)374-382
Number of pages9
JournalMolekulyarnaya Biologiya
Volume28
Issue number2
StatePublished - Jan 1 1994

ASJC Scopus subject areas

  • Medicine(all)

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