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Title: Alanine Scanning Mutagenesis Identifies an Asparagine–Arginine–Lysine Triad Essential to Assembly of the Shell of the Pdu Microcompartment

Abstract

Bacterial microcompartments (MCPs) are the simplest organelles known. They function to enhance metabolic pathways by confining several related enzymes inside an all-protein envelope called the shell. In this study, we investigated the factors that govern MCP assembly by performing scanning mutagenesis on the surface residues of PduA, a major shell protein of the MCP used for 1,2-propanediol degradation. Biochemical, genetic, and structural analysis of 20 mutants allowed us to determine that PduA K26, N29, and R79 are crucial residues that stabilize the shell of the 1,2-propanediol MCP. In addition, we identify two PduA mutants (K37A and K55A) that impair MCP function most likely by altering the permeability of its protein shell. These are the first studies to examine the phenotypic effects of shell protein structural mutations in an MCP system. The findings reported here may be applicable to engineering protein containers with improved stability for biotechnology applications.

Authors:
; ; ; ; ; ;
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
National Institutes of Health (NIH)
OSTI Identifier:
1132645
Resource Type:
Journal Article
Journal Name:
Journal of Molecular Biology
Additional Journal Information:
Journal Volume: 426; Journal Issue: 12; Journal ID: ISSN 0022-2836
Publisher:
Elsevier
Country of Publication:
United States
Language:
ENGLISH

Citation Formats

Sinha, Sharmistha, Cheng, Shouqiang, Sung, Yea Won, McNamara, Dan E., Sawaya, Michael R., Yeates, Todd O., and Bobik, Thomas A. Alanine Scanning Mutagenesis Identifies an Asparagine–Arginine–Lysine Triad Essential to Assembly of the Shell of the Pdu Microcompartment. United States: N. p., 2014. Web. doi:10.1016/j.jmb.2014.04.012.
Sinha, Sharmistha, Cheng, Shouqiang, Sung, Yea Won, McNamara, Dan E., Sawaya, Michael R., Yeates, Todd O., & Bobik, Thomas A. Alanine Scanning Mutagenesis Identifies an Asparagine–Arginine–Lysine Triad Essential to Assembly of the Shell of the Pdu Microcompartment. United States. https://doi.org/10.1016/j.jmb.2014.04.012
Sinha, Sharmistha, Cheng, Shouqiang, Sung, Yea Won, McNamara, Dan E., Sawaya, Michael R., Yeates, Todd O., and Bobik, Thomas A. 2014. "Alanine Scanning Mutagenesis Identifies an Asparagine–Arginine–Lysine Triad Essential to Assembly of the Shell of the Pdu Microcompartment". United States. https://doi.org/10.1016/j.jmb.2014.04.012.
@article{osti_1132645,
title = {Alanine Scanning Mutagenesis Identifies an Asparagine–Arginine–Lysine Triad Essential to Assembly of the Shell of the Pdu Microcompartment},
author = {Sinha, Sharmistha and Cheng, Shouqiang and Sung, Yea Won and McNamara, Dan E. and Sawaya, Michael R. and Yeates, Todd O. and Bobik, Thomas A.},
abstractNote = {Bacterial microcompartments (MCPs) are the simplest organelles known. They function to enhance metabolic pathways by confining several related enzymes inside an all-protein envelope called the shell. In this study, we investigated the factors that govern MCP assembly by performing scanning mutagenesis on the surface residues of PduA, a major shell protein of the MCP used for 1,2-propanediol degradation. Biochemical, genetic, and structural analysis of 20 mutants allowed us to determine that PduA K26, N29, and R79 are crucial residues that stabilize the shell of the 1,2-propanediol MCP. In addition, we identify two PduA mutants (K37A and K55A) that impair MCP function most likely by altering the permeability of its protein shell. These are the first studies to examine the phenotypic effects of shell protein structural mutations in an MCP system. The findings reported here may be applicable to engineering protein containers with improved stability for biotechnology applications.},
doi = {10.1016/j.jmb.2014.04.012},
url = {https://www.osti.gov/biblio/1132645}, journal = {Journal of Molecular Biology},
issn = {0022-2836},
number = 12,
volume = 426,
place = {United States},
year = {Sun Jun 01 00:00:00 EDT 2014},
month = {Sun Jun 01 00:00:00 EDT 2014}
}