Substrate binding to BamD triggers a conformational change in BamA to control membrane insertion

James Lee; Holly A. Sutterlin; Joseph S. Wzorek; Michael D. Mandler; Christine L. Hagan; Marcin Grabowicz; David Tomasek; Mary D. May; Elizabeth M. Hart; Thomas J. Silhavy; Daniel Kahne

doi:10.1073/pnas.1711727115

Substrate binding to BamD triggers a conformational change in BamA to control membrane insertion

James Lee, Holly A. Sutterlin, Joseph S. Wzorek, Michael D. Mandler, Christine L. Hagan, Marcin Grabowicz, David Tomasek, Mary D. May, Elizabeth M. Hart, Thomas J. Silhavy, Daniel Kahne

Molecular Biology

Research output: Contribution to journal › Article › peer-review

45 Scopus citations

Abstract

The β-barrel assembly machine (Bam) complex folds and inserts integral membrane proteins into the outer membrane of Gram-negative bacteria. The two essential components of the complex, BamA and BamD, both interact with substrates, but how the two coordinate with each other during assembly is not clear. To elucidate aspects of this process we slowed the assembly of an essential β-barrel substrate of the Bam complex, LptD, by changing a conserved residue near the C terminus. This defective substrate is recruited to the Bam complex via BamD but is unable to integrate into the membrane efficiently. Changes in the extracellular loops of BamA partially restore assembly kinetics, implying that BamA fails to engage this defective substrate. We conclude that substrate binding to BamD activates BamA by regulating extracellular loop interactions for folding and membrane integration.

Original language	English (US)
Pages (from-to)	2359-2364
Number of pages	6
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	115
Issue number	10
DOIs	https://doi.org/10.1073/pnas.1711727115
State	Published - Mar 6 2018

All Science Journal Classification (ASJC) codes

General

Keywords

Bam complex
Beta-barrel
Outer membrane
Protein folding

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10.1073/pnas.1711727115

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Lee, J., Sutterlin, H. A., Wzorek, J. S., Mandler, M. D., Hagan, C. L., Grabowicz, M., Tomasek, D., May, M. D., Hart, E. M., Silhavy, T. J., & Kahne, D. (2018). Substrate binding to BamD triggers a conformational change in BamA to control membrane insertion. Proceedings of the National Academy of Sciences of the United States of America, 115(10), 2359-2364. https://doi.org/10.1073/pnas.1711727115

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title = "Substrate binding to BamD triggers a conformational change in BamA to control membrane insertion",

abstract = "The β-barrel assembly machine (Bam) complex folds and inserts integral membrane proteins into the outer membrane of Gram-negative bacteria. The two essential components of the complex, BamA and BamD, both interact with substrates, but how the two coordinate with each other during assembly is not clear. To elucidate aspects of this process we slowed the assembly of an essential β-barrel substrate of the Bam complex, LptD, by changing a conserved residue near the C terminus. This defective substrate is recruited to the Bam complex via BamD but is unable to integrate into the membrane efficiently. Changes in the extracellular loops of BamA partially restore assembly kinetics, implying that BamA fails to engage this defective substrate. We conclude that substrate binding to BamD activates BamA by regulating extracellular loop interactions for folding and membrane integration.",

keywords = "Bam complex, Beta-barrel, Outer membrane, Protein folding",

author = "James Lee and Sutterlin, {Holly A.} and Wzorek, {Joseph S.} and Mandler, {Michael D.} and Hagan, {Christine L.} and Marcin Grabowicz and David Tomasek and May, {Mary D.} and Hart, {Elizabeth M.} and Silhavy, {Thomas J.} and Daniel Kahne",

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doi = "10.1073/pnas.1711727115",

language = "English (US)",

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Lee, J, Sutterlin, HA, Wzorek, JS, Mandler, MD, Hagan, CL, Grabowicz, M, Tomasek, D, May, MD, Hart, EM, Silhavy, TJ & Kahne, D 2018, 'Substrate binding to BamD triggers a conformational change in BamA to control membrane insertion', Proceedings of the National Academy of Sciences of the United States of America, vol. 115, no. 10, pp. 2359-2364. https://doi.org/10.1073/pnas.1711727115

TY - JOUR

T1 - Substrate binding to BamD triggers a conformational change in BamA to control membrane insertion

AU - Lee, James

AU - Sutterlin, Holly A.

AU - Wzorek, Joseph S.

AU - Mandler, Michael D.

AU - Hagan, Christine L.

AU - Grabowicz, Marcin

AU - Tomasek, David

AU - May, Mary D.

AU - Hart, Elizabeth M.

AU - Silhavy, Thomas J.

AU - Kahne, Daniel

PY - 2018/3/6

Y1 - 2018/3/6

N2 - The β-barrel assembly machine (Bam) complex folds and inserts integral membrane proteins into the outer membrane of Gram-negative bacteria. The two essential components of the complex, BamA and BamD, both interact with substrates, but how the two coordinate with each other during assembly is not clear. To elucidate aspects of this process we slowed the assembly of an essential β-barrel substrate of the Bam complex, LptD, by changing a conserved residue near the C terminus. This defective substrate is recruited to the Bam complex via BamD but is unable to integrate into the membrane efficiently. Changes in the extracellular loops of BamA partially restore assembly kinetics, implying that BamA fails to engage this defective substrate. We conclude that substrate binding to BamD activates BamA by regulating extracellular loop interactions for folding and membrane integration.

AB - The β-barrel assembly machine (Bam) complex folds and inserts integral membrane proteins into the outer membrane of Gram-negative bacteria. The two essential components of the complex, BamA and BamD, both interact with substrates, but how the two coordinate with each other during assembly is not clear. To elucidate aspects of this process we slowed the assembly of an essential β-barrel substrate of the Bam complex, LptD, by changing a conserved residue near the C terminus. This defective substrate is recruited to the Bam complex via BamD but is unable to integrate into the membrane efficiently. Changes in the extracellular loops of BamA partially restore assembly kinetics, implying that BamA fails to engage this defective substrate. We conclude that substrate binding to BamD activates BamA by regulating extracellular loop interactions for folding and membrane integration.

KW - Bam complex

KW - Beta-barrel

KW - Outer membrane

KW - Protein folding

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JF - Proceedings of the National Academy of Sciences of the United States of America

IS - 10

ER -

Substrate binding to BamD triggers a conformational change in BamA to control membrane insertion

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