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. 1975 Apr;72(4):1254–1257. doi: 10.1073/pnas.72.4.1254

Molecular basis of beta-galactosidase alpha-complementation.

K E Langley, M R Villarejo, A V Fowler, P J Zamenhof, I Zabin
PMCID: PMC432510  PMID: 1093175

Abstract

In previous studies, a cyanogen bromide peptide derived from amino-acid residues 3-92 of beta-galactosidase (EC 3.2.1.23; beta-D-galactoside galactohydrolase) was shown to have alpha-donor activity in intracistronic alpha-complementation. We have now isolated the defective beta-galactosidase alpha-acceptor protein from the deletion mutant strain M15 of Escherichia coli and find that it lacks residues 11-41 of betal-galactosidase. This is demonstrated by the isolation and sequence determination of a cyanogen bromide peptide from the M15 protein, which is identical to the corresponding peptide from beta-galactosidase except for the missing amino acids. We conclude that the alpha-donor peptide restores the region missing in the M15 protein.

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Selected References

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

  1. ADELBERG E. A., BURNS S. N. Genetic variation in the sex factor of Escherichia coli. J Bacteriol. 1960 Mar;79:321–330. doi: 10.1128/jb.79.3.321-330.1960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anfinsen C. B. Principles that govern the folding of protein chains. Science. 1973 Jul 20;181(4096):223–230. doi: 10.1126/science.181.4096.223. [DOI] [PubMed] [Google Scholar]
  3. BECKWITH J. R. A DELETION ANALYSIS OF THE LAC OPERATOR REGION IN ESCHERICHIA COLI. J Mol Biol. 1964 Mar;8:427–430. doi: 10.1016/s0022-2836(64)80206-0. [DOI] [PubMed] [Google Scholar]
  4. Bruton C. J., Hartley B. S. Chemical studies on methionyl-tRNA synthetase from Escherichia coli. J Mol Biol. 1970 Sep 14;52(2):165–178. doi: 10.1016/0022-2836(70)90023-9. [DOI] [PubMed] [Google Scholar]
  5. CRESTFIELD A. M., MOORE S., STEIN W. H. The preparation and enzymatic hydrolysis of reduced and S-carboxymethylated proteins. J Biol Chem. 1963 Feb;238:622–627. [PubMed] [Google Scholar]
  6. Fowler A. V. High-level production of -galactosidase by Escherichia coli merodiploids. J Bacteriol. 1972 Nov;112(2):856–860. doi: 10.1128/jb.112.2.856-860.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fowler A. V. The amino acid sequence of -galactosidase. II. Tryptic peptides of the maleylated protein and sequences of some tryptic peptides. J Biol Chem. 1972 Sep 10;247(17):5425–5431. [PubMed] [Google Scholar]
  8. Fowler A. V., Zabin I. Co-linearity of beta-galactosidase with its gene by immunological detection of incomplete polypeptide chains. Science. 1966 Nov 25;154(3752):1027–1029. doi: 10.1126/science.154.3752.1027. [DOI] [PubMed] [Google Scholar]
  9. Fowler A. V., Zabin I. The amino acid sequence of beta galactosidase. I. Isolation and composition of tryptic peptides. J Biol Chem. 1970 Oct 10;245(19):5032–5041. [PubMed] [Google Scholar]
  10. Goldberg M. E. Tertiary structure of Escherichia coli beta-D-galactosidase. J Mol Biol. 1969 Dec 28;46(3):441–446. doi: 10.1016/0022-2836(69)90187-9. [DOI] [PubMed] [Google Scholar]
  11. JACOB F., ULLMAN A., MONOD J. LE PROMOTEUR, 'EL'EMENT G'EN'ETIQUE N'ECESSAIRE 'A L'EXPRESSION D'UN OP'ERON. C R Hebd Seances Acad Sci. 1964 Mar 16;258:3125–3128. [PubMed] [Google Scholar]
  12. JOVIN T., CHRAMBACH A., NAUGHTON M. A. AN APPARATUS FOR PREPARATIVE TEMPERATURE-REGULATED POLYACRYLAMIDE GEL ELECTROPHORESIS. Anal Biochem. 1964 Nov;9:351–369. doi: 10.1016/0003-2697(64)90192-7. [DOI] [PubMed] [Google Scholar]
  13. Lin S., Villarejo M., Zabin I. Beta-galactosidase: alpha-complementation of a deletion mutant with cyanogen bromide peptides. Biochem Biophys Res Commun. 1970 Jul 27;40(2):249–254. doi: 10.1016/0006-291x(70)91002-8. [DOI] [PubMed] [Google Scholar]
  14. Lin S., Zabin I. Beta-galactosidase. Rates of synthesis and degradation of incomplete chains. J Biol Chem. 1972 Apr 10;247(7):2205–2211. [PubMed] [Google Scholar]
  15. Malamy M. H. Frameshift mutations in the lactose operon of E. coli. Cold Spring Harb Symp Quant Biol. 1966;31:189–201. doi: 10.1101/sqb.1966.031.01.027. [DOI] [PubMed] [Google Scholar]
  16. Morrison S. L., Zipser D. Polypeptide products of nonsense mutations. I. Termination fragments from nonsense mutations in the Z gene of the lac operon of Escherichia coli. J Mol Biol. 1970 Jun 14;50(2):359–371. doi: 10.1016/0022-2836(70)90198-1. [DOI] [PubMed] [Google Scholar]
  17. RICHARDS F. M., VITHAYATHIL P. J. The preparation of subtilisn-modified ribonuclease and the separation of the peptide and protein components. J Biol Chem. 1959 Jun;234(6):1459–1465. [PubMed] [Google Scholar]
  18. STEERS E., Jr, CRAVEN G. R., ANFINSEN C. B., BETHUNE J. L. EVIDENCE FOR NONIDENTICAL CHAINS IN THE BETA-GALACTOSIDASE OF ESCHERICHIA COLI K12. J Biol Chem. 1965 Jun;240:2478–2484. [PubMed] [Google Scholar]
  19. Steers E., Jr, Cuatrecasas P., Pollard H. B. The purification of beta-galactosidase from Escherichia coli by affinity chromatography. J Biol Chem. 1971 Jan 10;246(1):196–200. [PubMed] [Google Scholar]
  20. Ullmann A., Jacob F., Monod J. Characterization by in vitro complementation of a peptide corresponding to an operator-proximal segment of the beta-galactosidase structural gene of Escherichia coli. J Mol Biol. 1967 Mar 14;24(2):339–343. doi: 10.1016/0022-2836(67)90341-5. [DOI] [PubMed] [Google Scholar]
  21. Villarejo M. R., Zabin I. Affinity chromatography of -galactosidase fragments. Nat New Biol. 1973 Mar 14;242(115):50–52. doi: 10.1038/newbio242050a0. [DOI] [PubMed] [Google Scholar]
  22. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  23. Zabin I., Fowler A. V. The amino acid sequence of -galactosidase. 3. The sequences of NH 2 - and COOH-terminal tryptic peptides. J Biol Chem. 1972 Sep 10;247(17):5432–5435. [PubMed] [Google Scholar]
  24. Zamenhof P. J., Villarejo M. Construction and properties of Escherichia coli strains exhibiting -complementation of -galactosidase fragments in vivo. J Bacteriol. 1972 Apr;110(1):171–178. doi: 10.1128/jb.110.1.171-178.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]

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