1887

Abstract

A novel second streptomycete cyclophilin gene - designated - was isolated from a cosmid gene library of by using as gene probe a fragment of the previously isolated cyclophilin gene of the same organism. From its sequence the gene should encode a protein of 18868. Expression of in as a hexaHis-tagged fusion protein (H6ScCypB) and enzymic characterization of the purified protein showed that, like ScCypA, ScCypB is a peptidyl-prolyl isomerase (PPIase). The specific activity and substrate specificity of the enzyme were comparable to that of ScCypA, but it was threefold less sensitive to inhibition by cyclosporin A (CsA). In contrast to ScCypA, which is abundant and exists in free and liganded form, ScCypB was 50- to 100-fold less abundant in cytosol-derived protein fractions of or , as revealed by Western blot analyses, suggesting a specialized function for this enzyme in the streptomycete cell. Both and were found to be present as single copies in the genome of and hybridized to a single band in chromosomal DNAs of other streptomycetes. High-level expression of as well as of cloned into the expression vector pIJ702 did not produce detectable changes in growth and morphology of and Calculations of similarities to known cyclophilin sequences and construction of phylogenetic trees indicated that ScCypB and ScCypA are phylogenetically distant from each other. While ScCypA is clearly related to the eukaryotic cyclophilins, the analyses show the sequence of ScCypB to be the most divergent of all cyclophilin sequences, indicating that it possibly constitutes a cluster by itself.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-143-1-117
1997-01-01
2021-07-23
Loading full text...

Full text loading...

/deliver/fulltext/micro/143/1/mic-143-1-117.html?itemId=/content/journal/micro/10.1099/00221287-143-1-117&mimeType=html&fmt=ahah

References

  1. Bradford M.M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.. Anal Biochem 12:248–254
    [Google Scholar]
  2. Chen M.X., Chen Y.H., Cohen P.T.W. 1992; Polymerase chain reactions using Saccharomyces, Drosophila and human DNA predict a large family of protein serine/threonine phosphatases.. FEBS Lett 306:54–58
    [Google Scholar]
  3. Compton L.A., Davis J.M., MacDonald J.R., Bächinger H.P. 1992; Structural and functional characterization of Escherichia coli peptidylprolyl cis/trans isomerases.. Eur J Biochem 206:927–934
    [Google Scholar]
  4. Fischer G. 1994; Peptidyl-prolyl cis/trans isomerases and their effectors.. Angew Chem Int Ed 33:1415–1436
    [Google Scholar]
  5. Fischer G., Wittmann-Liebold B., Lang B.K., Kiefhaber T., Schmid F.X. 1989; Cyclophilin and peptidyl-prolyl cis-trans isomerase are probably identical proteins.. Nature 337:476–478
    [Google Scholar]
  6. Foor F., Parent S.A., Morin N., Dahl A.M., Ramadan N., Chrebet G., Bostian K.A., Nielsen J.B. 1992; Calcineurin mediates inhibition by FK506 and cyclosporin of recovery from alpha-factor arrest in yeast.. Nature 360:682–684
    [Google Scholar]
  7. Franco L., Jiminez A., Demolder J., Molemans F., Fiers W., Contreras R. 1991; The nucleotide sequence of a 3d cyclophilin-homologous gene from Saccharomyces cerevisiae.. Yeast 7:791–797
    [Google Scholar]
  8. Fruman D.A., Burakoff S.J., Bierer B. 1994; Immunophilins in protein folding and immunosuppression.. FASEB J 8:391–400
    [Google Scholar]
  9. Galat A. 1993; Peptidylproline cis-trans-isomerases: immunophilins.. Eur J Biochem 216:689–707
    [Google Scholar]
  10. Gasser C.S., Gunning D.A., Budelier K.A., Brown S.M. 1990; Structure and expression of cytosolic cyclophilin/peptidyl-prolyl cis/trans isomerase of higher plants and production of active tomato cyclophilin in Escherichia coli.. Proc Natl Acad Sci USA 879519–9523
    [Google Scholar]
  11. Gill S.C., von Hippel P.H. 1989; Calculation of protein extinction coefficients from amino acid sequence data.. Anal Biochem 182:319–326
    [Google Scholar]
  12. Handschumacher R.E., Harding M.W., Rice J., Drugge R.J. 1984; Cyclophilin: a specific cytosolic binding protein for cyclosporin A.. Science 226:544–546
    [Google Scholar]
  13. Harrison R.K., Stein R.L. 1990; Substrate specificities of the peptidyl prolyl cis/trans isomerase activities of cyclophilin and FK-506 binding protein: evidence for the existence of a family of distinct enzymes.. Biochemistry 29:3813–3816
    [Google Scholar]
  14. Hayano T., Takashashi N., Kato S., Maki N., Suzuki M. 1991; Two distinct forms of peptidylprolyl-cis-trans-isomerase are expressed separately in periplasmic and cytoplasmic compartments of Escherichia coli cells.. Biochemistry 30:3041–3048
    [Google Scholar]
  15. Herrler M., Bang H., Marahiel M.A. 1994; Cloning and characterization of ppib, a Bacillus subtilis gene which encodes a cyclosporin A-sensitive peptidyl-prolyl cis-trans isomerase.. Mol Microbiol 11:1073–1083
    [Google Scholar]
  16. High K.P. 1995; The antimicrobial activities of cyclosporine, FK506 and rapamycin.. Transplantation 57:1689–1700
    [Google Scholar]
  17. Higgins D.G., Bleasby A.J., Fuchs R. 1992; clustal v: improved software for multiple sequence alignment.. Comput Appl Biosci 8:189–191
    [Google Scholar]
  18. Hoffmann K., Kakalis L.T., Anderson K.S., Armitage I.M., Handschumacher R.E. 1995; Expression of human cyclophilin-40 and the effect of the His 141 Trp mutation on catalysis and cyclosporin A binding.. Eur J Biochem 229:188–193
    [Google Scholar]
  19. Hopwood D.A., Bibb M.J., Chater K.F., Kieser T., Bruton C.J., Kieser H.M., Lydiate D.J., Smith C.P., Ward J.M., Schrempf H. 1985 Genetic manipulations of Streptomyces Norwich: John Innes Foundation;
    [Google Scholar]
  20. Kallen J., Walkinshaw M.D. 1992; The X-ray structure of a tetrapeptide bound to the active site of human cyclophilin A.. FEBS Lett 300:286–290
    [Google Scholar]
  21. Kallen J., Spitzfaden C., Zurini M.G.M., Wider G., Widmer H., Wüthrich K., Walkinshaw M.D. 1991; Structure of human cyclophilin and its binding site for cyclosporin-A determined by X-ray crystallography and NMR spectroscopy.. Nature 353:276–279
    [Google Scholar]
  22. Katz E., Thompson C.J., Hopwood D.A. 1983; Cloning and expression of the tyrosinase gene from Streptomyces antibioticus in Streptomyces lividans.. J Gen Microbiol 129:2703–2714
    [Google Scholar]
  23. Keller U., Krengel U., Haese A. 1985; Genetic analysis in Streptomyces chrysomallus.. J Gen Microbiol 131:1181–1191
    [Google Scholar]
  24. Kieffer L.J., Thalhammer T., Handschumacher R.E. 1992; Isolation and characterization of a 40 kDa cyclophilin-related protein.. J Biol Chem 267:5503–5507
    [Google Scholar]
  25. Kofron J.L., Kuzmic P., Kishore V., Colón-Bonilla E., Rich D.H. 1991; Determination of kinetic constants for peptidyprolyl cis-trans isomerases by an improved spectrophotometric assay.. Biochemistry 30:6127–6134
    [Google Scholar]
  26. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4.. Nature 227:680–685
    [Google Scholar]
  27. Liu J., Chen C.-M., Walsh C. T. 1991a; Human and Escherichia coli cyclophilins: sensitivity to inhibition by the immuno-suppressant cyclosporin A correlates with a specific tryptophan residue.. Biochemistry 30:2306–2310
    [Google Scholar]
  28. Liu J., Farmer J. D. Jr Lane W. S., Friedman J., Weissman I., Schreiber S. L. 1991b; Calcineurin is a common target of cyclophilin-cyclosporin A and FKBP-FK506 complexes.. Cell 66:807–814
    [Google Scholar]
  29. Maniatis T., Fritsch E.F., Sambrook J. 1982 Molecular cloning. A Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  30. Mead D.A., Szczesn-Skorupa E., Kemper B. 1988; Singlestranded DNA blue T7 promoter plasmids: a versatile tandem promoter system for cloning and protein engineering.. Protein Eng I:67–74
    [Google Scholar]
  31. Page A.P., Landry D., Wilson G.G., Carlow K.S. 1995; Molecular characterization of a cyclosporin A-insensitive cyclo- philin from the parasitic nematode Brugia malayi.. Biochemistry 34:11545–11550
    [Google Scholar]
  32. Pahl A., Keller U. 1992; FK-506-binding proteins from streptomycetes producing immunosuppressive macrolactones of the FK-506 type.. J Bacteriol 174:5888–5894
    [Google Scholar]
  33. Pahl A., Ühlein M., Bang H., Schlumbohm W., Keller U. 1992; Streptomycetes possess peptidyl-prolyl cis-trans isomerases that strongly resemble cyclophilins from eukaryotic organisms.. Mol Microbiol 6:3551–3558
    [Google Scholar]
  34. Rahfeld J.U., Rucknagel K.P., Schelbert B., Ludwig B., Hacker J., Mann K., Fischer G. 1994; Confirmation of the existence of a third family among peptidyl-prolyl cis/trans isomerases - amino acid sequence and recombinant production of parvulin.. FEBS Lett 352:180–184
    [Google Scholar]
  35. Rinfret A., Collins C., Ménard R., Anderson S.K. 1994; The N-terminal cyclophilin-homologous domain of a 150-kilodalton tumor recognition molecule exhibits both peptidylprolyl cis-trans isomerase and chaperone activity.. Biochemistry 33:1668–1673
    [Google Scholar]
  36. Schönbrunner E.R., Mayer S., Tropschug M., Fischer G., Takahashi N., Schmid F. X. 1991; Catalysis of protein folding by cyclophilins from different species.. J Biol Chem 266:3630–3635
    [Google Scholar]
  37. Schreiber S.L., Crabtree G.R. 1992; The mechanism of action of cyclosporin-A and FK506.. Immunol Today 13:136–142
    [Google Scholar]
  38. Snyder S.H., Sabatini D.M. 1995; Immunophilins and the nervous system.. Nature Med 1:32–37
    [Google Scholar]
  39. Stamnes M.A., Shieh B.-H., Chuman L., Harris G.L., Zuker C.S. 1991; The cyclophilin homolog ninaA is a tissue-specific integral membrane protein required for the proper synthesis of a subset of Drosophila rhodopsins.. Cell 67:219–227
    [Google Scholar]
  40. Strohl W.R. 1992; Compilation and analysis of DNA sequences associated with apparent Streptomyces promoters.. Nucleic Acids Res 20:473–475
    [Google Scholar]
  41. Takahashi N., Hayano T., Suzuki M. 1989; Peptidyl-prolyl cis-trans isomerase is the cyclosporin A-binding protein cyclophilin.. Nature 337:473–475
    [Google Scholar]
  42. Theriault Y., Logan T.M., Meadows R., Yu L.P., Olejniczak E.T., Holzman T.F., Simmer R.L., Fesik S.W. 1993; Solution structure of the cyclosporin A cyclophilin complex by NMR.. Nature 361:88–91
    [Google Scholar]
  43. Trandinh C.C., Pao G.M., Saier M.H. 1992; Structural and evolutionary relationships among the immunophilins - 2 ubiquitous families of peptidyl-prolyl cis-trans isomerases.. FASEB J 6:3410–3420
    [Google Scholar]
  44. Tropschug M., Nicholson D., Hartl F., Kohler H., Pfanner N., Wachter E., Neupert W. 1988; Cyclosporin A-binding protein (cyclophilin) of Neurospora crassa.. J Biol Chem 263:14433–14440
    [Google Scholar]
  45. Xu Q., Leiva M.C, Fischkoff S.A., Handschumacher R.E., Lyttle CR. 1992; Leukocyte chemotactic activity of cyclophilin.. J Biol Chem 267:11968–11971
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-143-1-117
Loading
/content/journal/micro/10.1099/00221287-143-1-117
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error