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. 1994 May;3(5):730–736. doi: 10.1002/pro.5560030502

Determination of the binding frame within a physiological ligand for the chaperone SecB.

T B Topping 1, L L Randall 1
PMCID: PMC2142715  PMID: 8061603

Abstract

The hallmark of the class of proteins called chaperones is the amazing ability to bind tightly to a wide array of polypeptide ligands that have no consensus in sequence; chaperones recognize non-native structure. As a step in the elucidation of the molecular mechanism of such remarkable binding, we have characterized complexes between the bacterial chaperone SecB and a series of ligands related to maltose-binding protein. SecB interacts at multiple sites on its polypeptide ligand. The entire binding region covers approximately half of the primary sequence of maltose-binding protein and comprises contiguous sites positioned around the center of the sequence.

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Selected References

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  1. Breukink E., Kusters R., De Kruijff B. In-vitro studies on the folding characteristics of the Escherichia coli precursor protein prePhoE. Evidence that SecB prevents the precursor from aggregating by forming a functional complex. Eur J Biochem. 1992 Sep 1;208(2):419–425. doi: 10.1111/j.1432-1033.1992.tb17203.x. [DOI] [PubMed] [Google Scholar]
  2. Chun S. Y., Strobel S., Bassford P., Jr, Randall L. L. Folding of maltose-binding protein. Evidence for the identity of the rate-determining step in vivo and in vitro. J Biol Chem. 1993 Oct 5;268(28):20855–20862. [PubMed] [Google Scholar]
  3. Duplay P., Bedouelle H., Fowler A., Zabin I., Saurin W., Hofnung M. Sequences of the malE gene and of its product, the maltose-binding protein of Escherichia coli K12. J Biol Chem. 1984 Aug 25;259(16):10606–10613. [PubMed] [Google Scholar]
  4. Fikes J. D., Bassford P. J., Jr Export of unprocessed precursor maltose-binding protein to the periplasm of Escherichia coli cells. J Bacteriol. 1987 Jun;169(6):2352–2359. doi: 10.1128/jb.169.6.2352-2359.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Flynn G. C., Pohl J., Flocco M. T., Rothman J. E. Peptide-binding specificity of the molecular chaperone BiP. Nature. 1991 Oct 24;353(6346):726–730. doi: 10.1038/353726a0. [DOI] [PubMed] [Google Scholar]
  6. Gannon P. M., Li P., Kumamoto C. A. The mature portion of Escherichia coli maltose-binding protein (MBP) determines the dependence of MBP on SecB for export. J Bacteriol. 1989 Feb;171(2):813–818. doi: 10.1128/jb.171.2.813-818.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hardy S. J., Randall L. L. A kinetic partitioning model of selective binding of nonnative proteins by the bacterial chaperone SecB. Science. 1991 Jan 25;251(4992):439–443. doi: 10.1126/science.1989077. [DOI] [PubMed] [Google Scholar]
  8. Kumamoto C. A., Gannon P. M. Effects of Escherichia coli secB mutations on pre-maltose binding protein conformation and export kinetics. J Biol Chem. 1988 Aug 15;263(23):11554–11558. [PubMed] [Google Scholar]
  9. Landry S. J., Jordan R., McMacken R., Gierasch L. M. Different conformations for the same polypeptide bound to chaperones DnaK and GroEL. Nature. 1992 Jan 30;355(6359):455–457. doi: 10.1038/355455a0. [DOI] [PubMed] [Google Scholar]
  10. Langer T., Pfeifer G., Martin J., Baumeister W., Hartl F. U. Chaperonin-mediated protein folding: GroES binds to one end of the GroEL cylinder, which accommodates the protein substrate within its central cavity. EMBO J. 1992 Dec;11(13):4757–4765. doi: 10.1002/j.1460-2075.1992.tb05581.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lecker S., Lill R., Ziegelhoffer T., Georgopoulos C., Bassford P. J., Jr, Kumamoto C. A., Wickner W. Three pure chaperone proteins of Escherichia coli--SecB, trigger factor and GroEL--form soluble complexes with precursor proteins in vitro. EMBO J. 1989 Sep;8(9):2703–2709. doi: 10.1002/j.1460-2075.1989.tb08411.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Liu G. P., Topping T. B., Cover W. H., Randall L. L. Retardation of folding as a possible means of suppression of a mutation in the leader sequence of an exported protein. J Biol Chem. 1988 Oct 15;263(29):14790–14793. [PubMed] [Google Scholar]
  13. Liu G., Topping T. B., Randall L. L. Physiological role during export for the retardation of folding by the leader peptide of maltose-binding protein. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9213–9217. doi: 10.1073/pnas.86.23.9213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. MacIntyre S., Mutschler B., Henning U. Requirement of the SecB chaperone for export of a non-secretory polypeptide in Escherichia coli. Mol Gen Genet. 1991 Jun;227(2):224–228. doi: 10.1007/BF00259674. [DOI] [PubMed] [Google Scholar]
  15. Oliver D. B., Beckwith J. E. coli mutant pleiotropically defective in the export of secreted proteins. Cell. 1981 Sep;25(3):765–772. doi: 10.1016/0092-8674(81)90184-7. [DOI] [PubMed] [Google Scholar]
  16. Park K., Flynn G. C., Rothman J. E., Fasman G. D. Conformational change of chaperone Hsc70 upon binding to a decapeptide: a circular dichroism study. Protein Sci. 1993 Mar;2(3):325–330. doi: 10.1002/pro.5560020304. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Randall L. L., Hardy S. J. Correlation of competence for export with lack of tertiary structure of the mature species: a study in vivo of maltose-binding protein in E. coli. Cell. 1986 Sep 12;46(6):921–928. doi: 10.1016/0092-8674(86)90074-7. [DOI] [PubMed] [Google Scholar]
  18. Randall L. L. Peptide binding by chaperone SecB: implications for recognition of nonnative structure. Science. 1992 Jul 10;257(5067):241–245. doi: 10.1126/science.1631545. [DOI] [PubMed] [Google Scholar]
  19. Randall L. L., Topping T. B., Hardy S. J. No specific recognition of leader peptide by SecB, a chaperone involved in protein export. Science. 1990 May 18;248(4957):860–863. doi: 10.1126/science.2188362. [DOI] [PubMed] [Google Scholar]
  20. Schägger H., von Jagow G. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem. 1987 Nov 1;166(2):368–379. doi: 10.1016/0003-2697(87)90587-2. [DOI] [PubMed] [Google Scholar]
  21. Weiss J. B., Bassford P. J., Jr The folding properties of the Escherichia coli maltose-binding protein influence its interaction with SecB in vitro. J Bacteriol. 1990 Jun;172(6):3023–3029. doi: 10.1128/jb.172.6.3023-3029.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. de Cock H., Overeem W., Tommassen J. Biogenesis of outer membrane protein PhoE of Escherichia coli. Evidence for multiple SecB-binding sites in the mature portion of the PhoE protein. J Mol Biol. 1992 Mar 20;224(2):369–379. doi: 10.1016/0022-2836(92)91001-6. [DOI] [PubMed] [Google Scholar]

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