Summary
The gene encoding lysostaphin of Staphylococcus staphylolyticus was cloned in Escherichia coli and its DNA sequence was determined. The complete coding region comprises 1440 base pairs corresponding to a precursor of 480 amino acids (molecular weight 51669). It was shown by NH2-terminal amino acid sequence analysis of the purified extracellular lysostaphin from S. staphylolyticus that the mature lysostaphin consists of 246 amino acid residues (molecular weight 26926). Polyacrylamide gel electrophoresis revealed a similar molecular weight for the most active form. By computer analysis the secondary protein structure was predicted. It revealed three distinct regions in the precursor protein: a typical signal peptide (ca. 38 aa), a hydrophilic and highly ordered protein domain with 14 repetitive sequences (296 aa) and the hydrophobic mature lysostaphin. The lysostaphin precursor protein appears to be organized as a preprolysostaphin.
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Abbreviations
- aa:
-
amino acid(s)
References
Beachey EH, Seyer JM, Kang AH (1978) Repeating covalent structure of streptococal M protein. Proc Natl Acad Sci USA 75:3163–3167
Birnboim HL, Doly J (1979) A rapid alkaline extraction procedure for screening of recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523
Browder HP, Zygmunt WA, Young JR, Tavormina PA (1965) Lysostaphin: enzymatic mode of action. Biochem Biophys Res Commun 19:383–389
Chang JY, Brauer D, Wittmann-Liebold B (1978) Micro-sequence analysis of tides and proteins using 4-NN-dimethylaminoazobenzene 4′-isothiocyanate/phenylisothiocyanate double coupling method. FEBS Lett 93:205–214
Chen EY, Seeburg PH (1985) Supercoil sequencing: A fast and simple method for sequencing plasmid DNA. DNA 4:165–170
Chou PY, Fasman GD (1974) Prediction of protein conformation. Biochem 13:222–245
Clements S, Mehansho H, Carlson DM (1985) Novel multigene encoding highly repetitive peptide sequences. J Biol Chem 260:13471–13477
Cohen GH, Dietzschold B, Ponce de Leon M, Long D, Golub E, Varrichio A, Pereirs L, Eisenberg RJ (1984) Localization and synthesis of an antigenic determinant of herpes simplex virus glycoprotein D that stimulates the production of neutralizing antibody. J Virol 49:102–108
Garnier J, Osguthorpe DJ, Robson B (1978) Analysis of the accuracy and implication of simple methods for predicting the secondary structure of globular proteins. J Mol Biol 120:97–120
Götz F, Kreutz B, Schleifer KH (1983) Protoplast transformation of Staphylococcus carnosus by plasmid DNA. Mol Gen Genet 189:340–342
Götz F, Popp F, Korn E, Schleifer KH (1985) Complete nucleotide sequence of the lipase gene from Staphylococcus hyicus cloned in Staphylococcus carnosus. Nucleic Acids Res 13:5895–5906
Green CJ, Stewart GC, Hollis MA, Vold BS, Bott KF (1985) Nucleotide sequence of the Bacillus subtilis ribosomal RNA operon, rrnB. Gene 37:261–266
Hantke K, Braun V (1973) Covalent binding of lipid to protein: Diglyceride and amide-linked fatty acid at the N-terminal end of the murein-lipoprotein of the Escherichia coli outer membrane. Eur J Biochem 34:284–296
Heinrich P (1986) Guidelines for quick and simple plasmid sequencing. Handbook published by Boehringer Mannheim
Henikoff S (1984) Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene 28:351–359
Hennecke H, Günther I, Binder F (1982) A novel cloning vector for the direct selection of recombinant DNA in E. coli. Gene 19:231–234
Hopp TP, Woods KR (1981) Prediction of protein antigenic determinants amino acid sequences. Proc Natl Acad Sci USA 78:3824–3828
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Liebl W, Götz F (1986) Studies on lipse directed export of Escherichia coli β-lactamase in Staphylococcus carnosus Mol Gen Genet 204:166–173
Manjula BN, Fischetti VA (1980) Tropomyosin-like seven periodicity in three immunologically distinct streptococcal M proteins and its implications for the antiphagocytic properties of the molecule. J Exp Med 151:695–708
Matteuci MD, Caruthers MH (1980) The synthesis of oligodeoxy-pyrimidines on a polymer support. Tet Lett 21:719–722
Modrow S, Hahn BH, Shaw GM, Gallo RC, Wong-Staal F, Wolf H (1987) Computer-assisted analysis of envelope protein sequences of seven human immunodeficiency virus isolates: Prediction of antigenic epitopes in conserved and variable regions. J Virol 61:570–578
Neu HC, Heppel LA (1965) The release of enzymes from Escherichia coli by osmotic shock and during the formation of spheroplasts. J Biol Chem 240:3685–2692
Novick RP, Bouanchaud D (1971) Extrachromosomal nature of drug resistance in Staphylococcus aureus. Ann N Y Acad Sci 182:279–294
O'Callaghan CH, Morris A, Kriby SM, Shingler AH (1972) Novel method for detection of β-lactamase by using a chromogenic cephalosporin substrate. Antimicrob Agents Chemother 1:283–288
Perlman D, Halvorson HO (1983) A putative signal peptidase recognition site and sequence in eucaryotic and procaryotic signal peptides. J Mol Biol 167:391–409
Platt T (1986) Transcription termination and the regulation of gene expression. Annu Rev Biochem 55:339–372
Recsei PA, Gruss AD, Novick RP (1987) Cloning, sequence, and expression of the lysostaphin gene from Staphylococcus simulans. Proc Natl Acad Sci USA 84:1127–1131
Robinson JM, Hardman JK, Sloan GL (1979) Relationship between lysastaphin endopeptidase production and cell wall composition of Staphylococcus staphylolyticus. J Bacteriol 137:1158–1164
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Schindler CA, Schuhardt VT (1964) Lysostaphin: A new bacteriolytic agent for the Staphylococcus. Proc Natl Acad Sci USA 51:414–421
Schindler CA, Schuhardt VT (1965) Purification and properties of lysostaphin- a lytic agent for Staphylococcus aureus. Biochem Biophys Acta 97:242–250
Schleifer KH, Kandler O (1972) Peptidoglycan types of bacterial cell walls and their taxonomic implications. Bacteriol Rev 36:407–477
Schleifer KH, Kloos WE (1975) Isolation and characterization of staphylococci from human skin. I. Amended descriptions of Staphylococcus epidermis and Staphylococcus saprophyticus and descriptions of three new species: Staphylococcus cohnii, Staphylococcus haemolyticus, and Staphylococcus xylosus. Int J Syst Bacteriol 25:50–61
Shine J, Dalgarno L (1974) The 3′ terminal sequence of Escherichia coli ribosomal RNA: complementary to nonsense triplets and ribosome binding site. Proc Natl Acad Sci USA 71:1342–1346
Sjödahl J (1977) Repetitive sequences in protein A from Staphylococcus aureus. Eur J Biochem 73:343–351
Trayer HR, Buckley CE (1970) III: Molecular properties of lysostaphin, a bacteriolytic agent for Staphylococcus aureus. 245:4842–4846
Thudt K, Schleifer KH, Götz F (1985) Cloning and expression of alpha amylase gene from Bacillus stearothermophilus in Staphylococcus carnosus and other staphylococcal species. Gene 37:163–169
Uhlén M, Guss B, Nilsson B, Gatenbeck S, Philipson L, Lindberg M (1984) Complete sequence of the staphylococcal gene encoding protein A: A gene evolved through multiple duplications. J Biol Chem 259:1695–1702
Vieira J, Messing J (1982) The pUC plasmids, and M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19:259–268
Yanisch-Perron C, Vieira J, Messing J (1985) Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33:103–119
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Communicated by J. Lengeler
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Heinrich, P., Rosenstein, R., Böhmer, M. et al. The molecular organization of the lysostaphin gene and its sequences repeated in tandem. Mol Gen Genet 209, 563–569 (1987). https://doi.org/10.1007/BF00331163
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DOI: https://doi.org/10.1007/BF00331163