Secretion of Beta-lactamase into the Periplasm of Escherichia Coli: Evidence for a Distinct Release Step Associated with a Conformational Change

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1986 Jun 1

PMID 3520569

Citations 50

Authors

A MINSKY

R G Summers

J R Knowles

Affiliations

Soon will be listed here.

Abstract

The secretion of beta-lactamase (EC 3.5.2.6) into the periplasm of Escherichia coli has been followed by pulse-chase labeling at 15 degrees C. Though the periplasmic fraction contains only the mature form of the enzyme, the spheroplast fraction contains the completed precursor and a hitherto undocumented processed form. When whole spheroplasts are treated with trypsin, the processed form in this fraction is completely digested. This is in contrast to the native mature enzyme localized in the periplasm, which is trypsin resistant. The beta-lactamase is evidently processed after translocation to a trypsin-sensitive form that is transiently bound to the periplasmic face of the inner membrane. The release of this processed form into the periplasm occurs concomitantly with a conformational change that results in the soluble, catalytically active, trypsin-resistant structure.

Citing Articles

A High Throughput Assay for Measuring Secreted Protein Based on a Fluorescent Reporter Reveals Regulatory and Structural Insights in Type Three Secretion System.

Leach S, Summers J, Wen E, Tullman-Ercek D bioRxiv. 2025; .

PMID: 39868124 PMC: 11760704. DOI: 10.1101/2025.01.17.633628.

A broadly cross-reactive i-body to AMA1 potently inhibits blood and liver stages of Plasmodium parasites.

Angage D, Chmielewski J, Maddumage J, Hesping E, Caiazzo S, Lai K Nat Commun. 2024; 15(1):7206.

PMID: 39174515 PMC: 11341838. DOI: 10.1038/s41467-024-50770-7.

Dynamics and quantitative contribution of the aminoglycoside 6'--acetyltransferase type Ib to amikacin resistance.

dUdekem dAcoz O, Hue F, Ye T, Wang L, Leroux M, Rajngewerc L mSphere. 2024; 9(3):e0078923.

PMID: 38353533 PMC: 10964403. DOI: 10.1128/msphere.00789-23.

A unifying mechanism for protein transport through the core bacterial Sec machinery.

Allen W, Collinson I Open Biol. 2023; 13(8):230166.

PMID: 37643640 PMC: 10465204. DOI: 10.1098/rsob.230166.

The biogenesis of β-lactamase enzymes.

Kaderabkova N, Bharathwaj M, Furniss R, Gonzalez D, Palmer T, Mavridou D Microbiology (Reading). 2022; 168(8).

PMID: 35943884 PMC: 10235803. DOI: 10.1099/mic.0.001217.

References

Josefsson L, Randall L . Different exported proteins in E. coli show differences in the temporal mode of processing in vivo. Cell. 1981; 25(1):151-7. DOI: 10.1016/0092-8674(81)90239-7. View

Ito K, Beckwith J . Role of the mature protein sequence of maltose-binding protein in its secretion across the E. coli cytoplasmic membrane. Cell. 1981; 25(1):143-50. DOI: 10.1016/0092-8674(81)90238-5. View

Walter P, Blobel G . Translocation of proteins across the endoplasmic reticulum. II. Signal recognition protein (SRP) mediates the selective binding to microsomal membranes of in-vitro-assembled polysomes synthesizing secretory protein. J Cell Biol. 1981; 91(2 Pt 1):551-6. PMC: 2111991. DOI: 10.1083/jcb.91.2.551. View

Walter P, Blobel G . Translocation of proteins across the endoplasmic reticulum III. Signal recognition protein (SRP) causes signal sequence-dependent and site-specific arrest of chain elongation that is released by microsomal membranes. J Cell Biol. 1981; 91(2 Pt 1):557-61. PMC: 2111983. DOI: 10.1083/jcb.91.2.557. View

Koshland D, Botstein D . Evidence for posttranslational translocation of beta-lactamase across the bacterial inner membrane. Cell. 1982; 30(3):893-902. DOI: 10.1016/0092-8674(82)90294-x. View

Kadonaga J, Gautier A, Straus D, Charles A, Edge M, Knowles J . The role of the beta-lactamase signal sequence in the secretion of proteins by Escherichia coli. J Biol Chem. 1984; 259(4):2149-54. View

Lopilato J, Garwin J, Emr S, Silhavy T, Beckwith J . D-ribose metabolism in Escherichia coli K-12: genetics, regulation, and transport. J Bacteriol. 1984; 158(2):665-73. PMC: 215481. DOI: 10.1128/jb.158.2.665-673.1984. View

Randall L . Translocation of domains of nascent periplasmic proteins across the cytoplasmic membrane is independent of elongation. Cell. 1983; 33(1):231-40. DOI: 10.1016/0092-8674(83)90352-5. View

Walter P, Gilmore R, Blobel G . Protein translocation across the endoplasmic reticulum. Cell. 1984; 38(1):5-8. DOI: 10.1016/0092-8674(84)90520-8. View

10.

Randall L, Hardy S . Export of protein in bacteria. Microbiol Rev. 1984; 48(4):290-8. PMC: 373220. DOI: 10.1128/mr.48.4.290-298.1984. View

11.

Hengge R, Boos W . Defective secretion of maltose- and ribose-binding proteins caused by a truncated periplasmic protein in Escherichia coli. J Bacteriol. 1985; 162(3):972-8. PMC: 215870. DOI: 10.1128/jb.162.3.972-978.1985. View

12.

CITRI N, Zyk N . The interaction of penicillinase with penicillins. IV. Structural aspects of catalytic and non-catalytic interactions. Biochim Biophys Acta. 1965; 99(3):427-41. DOI: 10.1016/s0926-6593(65)80197-7. View

13.

Tenu J, Viratelle O, Garnier J, Yon J . pH dependence of the activity of beta-galactosidase from Escherichia coli. Eur J Biochem. 1971; 20(3):363-70. DOI: 10.1111/j.1432-1033.1971.tb01402.x. View

14.

Plaut B, Knowles J . pH-dependence of the triose phosphate isomerase reaction. Biochem J. 1972; 129(2):311-20. PMC: 1174081. DOI: 10.1042/bj1290311. View

15.

Blobel G, Dobberstein B . Transfer of proteins across membranes. I. Presence of proteolytically processed and unprocessed nascent immunoglobulin light chains on membrane-bound ribosomes of murine myeloma. J Cell Biol. 1975; 67(3):835-51. PMC: 2111658. DOI: 10.1083/jcb.67.3.835. View

16.

Witholt B, Boekhout M, Brock M, Kingma J, Heerikhuizen H, Leij L . An efficient and reproducible procedure for the formation of spheroplasts from variously grown Escherichia coli. Anal Biochem. 1976; 74(1):160-70. DOI: 10.1016/0003-2697(76)90320-1. View

17.

Boquet P, Silverman M, PAPPENHEIMER Jr A, Vernon W . Binding of triton X-100 to diphtheria toxin, crossreacting material 45, and their fragments. Proc Natl Acad Sci U S A. 1976; 73(12):4449-53. PMC: 431494. DOI: 10.1073/pnas.73.12.4449. View

18.

Wickner W . The assembly of proteins into biological membranes: The membrane trigger hypothesis. Annu Rev Biochem. 1979; 48:23-45. DOI: 10.1146/annurev.bi.48.070179.000323. View

19.

Blobel G . Intracellular protein topogenesis. Proc Natl Acad Sci U S A. 1980; 77(3):1496-500. PMC: 348522. DOI: 10.1073/pnas.77.3.1496. View

20.

OXENDER D, Anderson J, Daniels C, Landick R, Gunsalus R, Zurawski G . Amino-terminal sequence and processing of the precursor of the leucine-specific binding protein, and evidence for conformational differences between the precursor and the mature form. Proc Natl Acad Sci U S A. 1980; 77(4):2005-9. PMC: 348639. DOI: 10.1073/pnas.77.4.2005. View