Cotranslational Folding of Alkaline Phosphatase in the Periplasm of Escherichia Coli

Overview

Journal Protein Sci

Specialty Biochemistry

Date 2020 Aug 14

PMID 32790204

Citations 7

Authors

Rageia Elfageih

Alexandros Karyolaimos

Grant Kemp

Jan-Willem de Gier

Gunnar von Heijne

Renuka Kudva

Affiliations

Soon will be listed here.

Abstract

Cotranslational protein folding studies using Force Profile Analysis, a method where the SecM translational arrest peptide is used to detect folding-induced forces acting on the nascent polypeptide, have so far been limited mainly to small domains of cytosolic proteins that fold in close proximity to the translating ribosome. In this study, we investigate the cotranslational folding of the periplasmic, disulfide bond-containing Escherichia coli protein alkaline phosphatase (PhoA) in a wild-type strain background and a strain background devoid of the periplasmic thiol: disulfide interchange protein DsbA. We find that folding-induced forces can be transmitted via the nascent chain from the periplasm to the polypeptide transferase center in the ribosome, a distance of ~160 Å, and that PhoA appears to fold cotranslationally via at least two disulfide-stabilized folding intermediates. Thus, Force Profile Analysis can be used to study cotranslational folding of proteins in an extra-cytosolic compartment, like the periplasm.

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References

Papanikou E, Karamanou S, Baud C, Frank M, Sianidis G, Keramisanou D . Identification of the preprotein binding domain of SecA. J Biol Chem. 2005; 280(52):43209-17. DOI: 10.1074/jbc.M509990200. View

Kimura E, Akita M, Matsuyama S, Mizushima S . Determination of a region in SecA that interacts with presecretory proteins in Escherichia coli. J Biol Chem. 1991; 266(10):6600-6. View

Tian P, Steward A, Kudva R, Su T, Shilling P, Nickson A . Folding pathway of an Ig domain is conserved on and off the ribosome. Proc Natl Acad Sci U S A. 2018; 115(48):E11284-E11293. PMC: 6275497. DOI: 10.1073/pnas.1810523115. View

Akiyama Y, Ito K . Folding and assembly of bacterial alkaline phosphatase in vitro and in vivo. J Biol Chem. 1993; 268(11):8146-50. View

Kadokura H, Tian H, Zander T, Bardwell J, Beckwith J . Snapshots of DsbA in action: detection of proteins in the process of oxidative folding. Science. 2004; 303(5657):534-7. DOI: 10.1126/science.1091724. View

Huber D, Jamshad M, Hanmer R, Schibich D, Doring K, Marcomini I . SecA Cotranslationally Interacts with Nascent Substrate Proteins In Vivo. J Bacteriol. 2016; 199(2). PMC: 5198489. DOI: 10.1128/JB.00622-16. View

Keseler I, Mackie A, Santos-Zavaleta A, Billington R, Bonavides-Martinez C, Caspi R . The EcoCyc database: reflecting new knowledge about Escherichia coli K-12. Nucleic Acids Res. 2016; 45(D1):D543-D550. PMC: 5210515. DOI: 10.1093/nar/gkw1003. View

Denks K, Vogt A, Sachelaru I, Petriman N, Kudva R, Koch H . The Sec translocon mediated protein transport in prokaryotes and eukaryotes. Mol Membr Biol. 2014; 31(2-3):58-84. DOI: 10.3109/09687688.2014.907455. View

Watanabe M, Blobel G . High-affinity binding of Escherichia coli SecB to the signal sequence region of a presecretory protein. Proc Natl Acad Sci U S A. 1995; 92(22):10133-6. PMC: 40750. DOI: 10.1073/pnas.92.22.10133. View

10.

Ullers R, Luirink J, Harms N, Schwager F, Georgopoulos C, Genevaux P . SecB is a bona fide generalized chaperone in Escherichia coli. Proc Natl Acad Sci U S A. 2004; 101(20):7583-8. PMC: 419649. DOI: 10.1073/pnas.0402398101. View

11.

Marsden A, Hollins J, ONeill C, Ryzhov P, Higson S, Mendonca C . Investigating the Effect of Chain Connectivity on the Folding of a Beta-Sheet Protein On and Off the Ribosome. J Mol Biol. 2018; 430(24):5207-5216. PMC: 6288478. DOI: 10.1016/j.jmb.2018.10.011. View

12.

Osborne A, Clemons Jr W, Rapoport T . A large conformational change of the translocation ATPase SecA. Proc Natl Acad Sci U S A. 2004; 101(30):10937-42. PMC: 491988. DOI: 10.1073/pnas.0401742101. View

13.

Singh R, Kraft C, Jaiswal R, Sejwal K, Kasaragod V, Kuper J . Cryo-electron microscopic structure of SecA protein bound to the 70S ribosome. J Biol Chem. 2014; 289(10):7190-7199. PMC: 3945378. DOI: 10.1074/jbc.M113.506634. View

14.

Ismail N, Hedman R, Linden M, von Heijne G . Charge-driven dynamics of nascent-chain movement through the SecYEG translocon. Nat Struct Mol Biol. 2015; 22(2):145-149. PMC: 4338579. DOI: 10.1038/nsmb.2940. View

15.

Chatzi K, Sardis M, Economou A, Karamanou S . SecA-mediated targeting and translocation of secretory proteins. Biochim Biophys Acta. 2014; 1843(8):1466-74. DOI: 10.1016/j.bbamcr.2014.02.014. View

16.

Bardwell J, McGovern K, Beckwith J . Identification of a protein required for disulfide bond formation in vivo. Cell. 1991; 67(3):581-9. DOI: 10.1016/0092-8674(91)90532-4. View

17.

Lill R, Dowhan W, Wickner W . The ATPase activity of SecA is regulated by acidic phospholipids, SecY, and the leader and mature domains of precursor proteins. Cell. 1990; 60(2):271-80. DOI: 10.1016/0092-8674(90)90742-w. View

18.

Jamshad M, Knowles T, White S, Ward D, Mohammed F, Rahman K . The C-terminal tail of the bacterial translocation ATPase SecA modulates its activity. Elife. 2019; 8. PMC: 6620043. DOI: 10.7554/eLife.48385. View

19.

Thomason L, Costantino N, Court D . E. coli genome manipulation by P1 transduction. Curr Protoc Mol Biol. 2008; Chapter 1:1.17.1-1.17.8. DOI: 10.1002/0471142727.mb0117s79. View

20.

Smets D, Loos M, Karamanou S, Economou A . Protein Transport Across the Bacterial Plasma Membrane by the Sec Pathway. Protein J. 2019; 38(3):262-273. DOI: 10.1007/s10930-019-09841-8. View