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. 1978 Apr;134(1):287–294. doi: 10.1128/jb.134.1.287-294.1978

Escherichia coli mutants deficient in the production of alkaline phosphatase isozymes.

A Nakata, M Yamaguchi, K Izutani, M Amemura
PMCID: PMC222245  PMID: 348683

Abstract

Escherichia coli K-12 mutants showing an altered isozyme pattern of alkaline phosphatase were isolated. Whereas wild-type strains synthesized all three isozymes in a synthetic medium supplemented with Casamino Acids or arginine but synthesized only isozyme 3 in a medium without supplement, the mutant strains synthesized isozyme 1 and a small amount (if any) of isozyme 2, but no isozyme 3, under all growth conditions. The mutation responsible for the altered isozyme pattern, designated iap, was mapped by P1 transduction in the interval between cysC and srl (at about 58.5 min on the E. coli genetic map). It was cotransducible with cysC and srl at frequencies of 0.54 and 0.08, respectively. The order of the genes in this region was srl-iap-cysC-argA-thyA-lysA. Three more independent mutations were also mapped in the same locus. We purified isozymes 1' and 3' from iap and iap+ strains and analyzed the sequences of four amino acids from the amino terminus of each polypeptide. They were Arg-Thr-Pro-Glu (or Gln) in isozyme 1' and Thr-Pro-Glu (or gln)-Met in isozyme 3', which were identical with those of corresponding isozymes produced by the wild-type phoA+ strain (P.M. Kelley, P.A. Neumann, K. Schriefer, F. Cancedda, M.J. Schlesinger, and R.A. Bradshaw, Biochemistry 12:3499-3503, 1973; M.J. Schlesinger, W. Bloch, and P.M. Kelley, p. 333-342, in Isozymes, Academic Press Inc., 1975). These results indicate that the different mobilities of isozymes 1, 2, and 3 are determined by the presence or absence of amino-terminal arginine residues in polypeptides.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bachmann B. J., Low K. B., Taylor A. L. Recalibrated linkage map of Escherichia coli K-12. Bacteriol Rev. 1976 Mar;40(1):116–167. doi: 10.1128/br.40.1.116-167.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bridgen J., Secher D. S. Molecular heterogeneity of alkaline phosphatase. FEBS Lett. 1973 Jan 1;29(1):55–57. doi: 10.1016/0014-5793(73)80014-6. [DOI] [PubMed] [Google Scholar]
  3. Csopak H., Garellick G., Hallberg B. Purification of Escherichia coli alkaline phosphatase. Improved growth conditions for the bacteria, modified methods of preparation of the enzyme. Acta Chem Scand. 1972;26(6):2401–2411. doi: 10.3891/acta.chem.scand.26-2401. [DOI] [PubMed] [Google Scholar]
  4. Edman P. Sequence determination. Mol Biol Biochem Biophys. 1970;8:211–255. doi: 10.1007/978-3-662-12834-3_8. [DOI] [PubMed] [Google Scholar]
  5. GAREN A., GAREN S. Complementation in vivo between structural mutants of alkaline phosphatase from E. coli. J Mol Biol. 1963 Jul;7:13–22. doi: 10.1016/s0022-2836(63)80015-7. [DOI] [PubMed] [Google Scholar]
  6. GAREN A., LEVINTHAL C. A fine-structure genetic and chemical study of the enzyme alkaline phosphatase of E. coli. I. Purification and characterization of alkaline phosphatase. Biochim Biophys Acta. 1960 Mar 11;38:470–483. doi: 10.1016/0006-3002(60)91282-8. [DOI] [PubMed] [Google Scholar]
  7. Kelley P. M., Neumann P. A., Shriefer K., Cancedda F., Schlesinger M. J., Bradshaw R. A. Amino acid sequence of Escherichia coli alkaline phosphatase. Amino- and carboxyl-terminal sequences and variations between two isozymes. Biochemistry. 1973 Aug 28;12(18):3499–3503. doi: 10.1021/bi00742a023. [DOI] [PubMed] [Google Scholar]
  8. Kulbe K. D. Micropolyamide thin-layer chromatography of phenylthiohydantoin amino acids (PTH) at subnanomolar level. A rapid microtechnique for simultaneous multisample identification after automated Edman degradations. Anal Biochem. 1974 Jun;59(2):564–573. doi: 10.1016/0003-2697(74)90310-8. [DOI] [PubMed] [Google Scholar]
  9. LEVINTHAL C., SIGNER E. R., FETHEROLF K. Reactivation and hybridization of reduced alkaline phosphatase. Proc Natl Acad Sci U S A. 1962 Jul 15;48:1230–1237. doi: 10.1073/pnas.48.7.1230. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lazdunski C., Lazdunski M. Les isophosphatases alcalines d' Escherichia coli. Séparation, propriétés cinétiques et structurales. Biochim Biophys Acta. 1967 Oct 23;147(2):280–288. [PubMed] [Google Scholar]
  11. MALAMY M. H., HORECKER B. L. PURIFICATION AND CRYSTALLIZATION OF THE ALKALINE PHOSPHATASE OF ESCHERICHIA COLI. Biochemistry. 1964 Dec;3:1893–1897. doi: 10.1021/bi00900a018. [DOI] [PubMed] [Google Scholar]
  12. Nakata A., Amemura M., Yamaguchi M., Izutani K. Factors affecting the formation of alkaline phosphatase isozymes in Escherichia coli K-12. Biken J. 1977 Jun;20(2):47–55. [PubMed] [Google Scholar]
  13. Nakata A., Peterson G. R., Brooks E. L., Rothman F. G. Location and orientation of the phoA locus on the Escherichia coli K-12 linkage map. J Bacteriol. 1971 Sep;107(3):683–689. doi: 10.1128/jb.107.3.683-689.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Natori S., Garen A. Molecular heterogeneity in the amino-terminal region of alkaline phosphatase. J Mol Biol. 1970 May 14;49(3):577–588. doi: 10.1016/0022-2836(70)90282-2. [DOI] [PubMed] [Google Scholar]
  15. Piggot P. J., Sklar M. D., Gorini L. Ribosomal alterations controlling alkaline phosphatase isozymes in Escherichia coli. J Bacteriol. 1972 Apr;110(1):291–299. doi: 10.1128/jb.110.1.291-299.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. ROTHMAN F., BYRNE R. Fingerprint analysis of alkaline phosphatase of Escherichia coli K12. J Mol Biol. 1963 Apr;6:330–340. doi: 10.1016/s0022-2836(63)80092-3. [DOI] [PubMed] [Google Scholar]
  17. SIGNER E. R., TORRIANI A., LEVINTHAL C. Gene expression in intergeneric merozygotes. Cold Spring Harb Symp Quant Biol. 1961;26:31–34. doi: 10.1101/sqb.1961.026.01.008. [DOI] [PubMed] [Google Scholar]
  18. Schlesinger M. J., Andersen L. Multiple molecular forms of the alkaline phosphatase of Escherichia coli. Ann N Y Acad Sci. 1968 Jun 14;151(1):159–170. doi: 10.1111/j.1749-6632.1968.tb11886.x. [DOI] [PubMed] [Google Scholar]
  19. Suzuki T., Garen A. Fragments of alkaline phosphatase from nonsense mutants. I. Isolation and characterization of fragments from amber and ochre mutants. J Mol Biol. 1969 Nov 14;45(3):549–566. doi: 10.1016/0022-2836(69)90312-x. [DOI] [PubMed] [Google Scholar]
  20. Weiner A. M., Platt T., Weber K. Amino-terminal sequence analysis of proteins purified on a nanomole scale by gel electrophoresis. J Biol Chem. 1972 May 25;247(10):3242–3251. [PubMed] [Google Scholar]

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