Tags

Type your tag names separated by a space and hit enter

Crystal structures of the transpeptidase domain of the Mycobacterium tuberculosis penicillin-binding protein PonA1 reveal potential mechanisms of antibiotic resistance.
FEBS J. 2016 06; 283(12):2206-18.FJ

Abstract

Mycobacterium tuberculosis is a human respiratory pathogen that causes the deadly disease tuberculosis. The rapid global spread of antibiotic-resistant M. tuberculosis makes tuberculosis infections difficult to treat. To overcome this problem new effective antimicrobial strategies are urgently needed. One promising target for new therapeutic approaches is PonA1, a class A penicillin-binding protein, which is required for maintaining physiological cell wall synthesis and cell shape during growth in mycobacteria. Here, crystal structures of the transpeptidase domain, the enzymatic domain responsible for penicillin binding, of PonA1 from M. tuberculosis in the inhibitor-free form and in complex with penicillin V are reported. We used site-directed mutagenesis, antibiotic profiling experiments, and fluorescence thermal shift assays to measure PonA1's sensitivity to different classes of β-lactams. Structural comparison of the PonA1 apo-form and the antibiotic-bound form shows that binding of penicillin V induces conformational changes in the position of the loop β4'-α3 surrounding the penicillin-binding site. We have also found that binding of different antibiotics including penicillin V positively impacts protein stability, while other tested β-lactams such as clavulanate or meropenem resulted in destabilization of PonA1. Our antibiotic profiling experiments indicate that the transpeptidase activity of PonA1 in both M. tuberculosis and M. smegmatis mediates tolerance to specific cell wall-targeting antibiotics, particularly to penicillin V and meropenem. Because M. tuberculosis is an important human pathogen, these structural data provide a template to design novel transpeptidase inhibitors to treat tuberculosis infections.

DATABASE

Structural data are available in the PDB database under the accession numbers 5CRF and 5CXW.

Links

PMC Free PDF

Authors+Show Affiliations

Filippova EV
Department of Biochemistry and Molecular Genetics, Feinberg School of Medicine, Northwestern University, Chicago, IL, USA. Midwest Center for Structural Genomics (MCSG), Biosciences Division, Argonne National Laboratory, Argonne, IL, USA. Center for Structural Genomics of Infectious Diseases (CSGID), Feinberg School of Medicine, Northwestern University, Chicago, IL, USA.
Kieser KJ
Department of Immunology and Infectious Disease, Harvard T.H. Chan School of Public Health, Boston, MA, USA.
Luan CH
Midwest Center for Structural Genomics (MCSG), Biosciences Division, Argonne National Laboratory, Argonne, IL, USA. High Throughput Analysis Laboratory and Department of Molecular Biosciences, Northwestern University, Evanston, IL, USA.
Wawrzak Z
Life Science Collaborative Access Team, Synchrotron Research Center, Northwestern University, Evanston, IL, USA.
Kiryukhina O
Department of Biochemistry and Molecular Genetics, Feinberg School of Medicine, Northwestern University, Chicago, IL, USA. Midwest Center for Structural Genomics (MCSG), Biosciences Division, Argonne National Laboratory, Argonne, IL, USA. Center for Structural Genomics of Infectious Diseases (CSGID), Feinberg School of Medicine, Northwestern University, Chicago, IL, USA.
Rubin EJ
Department of Immunology and Infectious Disease, Harvard T.H. Chan School of Public Health, Boston, MA, USA. Department of Microbiology and Immunobiology, Harvard Medical School, Boston, MA, USA.
Anderson WF
Department of Biochemistry and Molecular Genetics, Feinberg School of Medicine, Northwestern University, Chicago, IL, USA. Midwest Center for Structural Genomics (MCSG), Biosciences Division, Argonne National Laboratory, Argonne, IL, USA. Center for Structural Genomics of Infectious Diseases (CSGID), Feinberg School of Medicine, Northwestern University, Chicago, IL, USA.

MeSH

Binding SitesCrystallography, X-RayDrug Resistance, MicrobialHumansMutagenesis, Site-DirectedMycobacterium tuberculosisPenicillin VPenicillin-Binding ProteinsPeptidyl TransferasesTuberculosisbeta-Lactams

Pub Type(s)

Editorial
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Research Support, N.I.H., Extramural

Language

eng

PubMed ID

27101811

Citation

Filippova, Ekaterina V., et al. "Crystal Structures of the Transpeptidase Domain of the Mycobacterium Tuberculosis Penicillin-binding Protein PonA1 Reveal Potential Mechanisms of Antibiotic Resistance." The FEBS Journal, vol. 283, no. 12, 2016, pp. 2206-18.
Filippova EV, Kieser KJ, Luan CH, et al. Crystal structures of the transpeptidase domain of the Mycobacterium tuberculosis penicillin-binding protein PonA1 reveal potential mechanisms of antibiotic resistance. FEBS J. 2016;283(12):2206-18.
Filippova, E. V., Kieser, K. J., Luan, C. H., Wawrzak, Z., Kiryukhina, O., Rubin, E. J., & Anderson, W. F. (2016). Crystal structures of the transpeptidase domain of the Mycobacterium tuberculosis penicillin-binding protein PonA1 reveal potential mechanisms of antibiotic resistance. The FEBS Journal, 283(12), 2206-18. https://doi.org/10.1111/febs.13738
Filippova EV, et al. Crystal Structures of the Transpeptidase Domain of the Mycobacterium Tuberculosis Penicillin-binding Protein PonA1 Reveal Potential Mechanisms of Antibiotic Resistance. FEBS J. 2016;283(12):2206-18. PubMed PMID: 27101811.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Crystal structures of the transpeptidase domain of the Mycobacterium tuberculosis penicillin-binding protein PonA1 reveal potential mechanisms of antibiotic resistance. AU - Filippova,Ekaterina V, AU - Kieser,Karen J, AU - Luan,Chi-Hao, AU - Wawrzak,Zdzislaw, AU - Kiryukhina,Olga, AU - Rubin,Eric J, AU - Anderson,Wayne F, Y1 - 2016/05/25/ PY - 2016/01/28/received PY - 2016/03/30/revised PY - 2016/04/15/accepted PY - 2016/4/23/entrez PY - 2016/4/23/pubmed PY - 2017/6/22/medline KW - PBP KW - PonA1 KW - antibiotic resistance KW - transpeptidase domain KW - tuberculosis KW - β-lactams SP - 2206 EP - 18 JF - The FEBS journal JO - FEBS J VL - 283 IS - 12 N2 - UNLABELLED: Mycobacterium tuberculosis is a human respiratory pathogen that causes the deadly disease tuberculosis. The rapid global spread of antibiotic-resistant M. tuberculosis makes tuberculosis infections difficult to treat. To overcome this problem new effective antimicrobial strategies are urgently needed. One promising target for new therapeutic approaches is PonA1, a class A penicillin-binding protein, which is required for maintaining physiological cell wall synthesis and cell shape during growth in mycobacteria. Here, crystal structures of the transpeptidase domain, the enzymatic domain responsible for penicillin binding, of PonA1 from M. tuberculosis in the inhibitor-free form and in complex with penicillin V are reported. We used site-directed mutagenesis, antibiotic profiling experiments, and fluorescence thermal shift assays to measure PonA1's sensitivity to different classes of β-lactams. Structural comparison of the PonA1 apo-form and the antibiotic-bound form shows that binding of penicillin V induces conformational changes in the position of the loop β4'-α3 surrounding the penicillin-binding site. We have also found that binding of different antibiotics including penicillin V positively impacts protein stability, while other tested β-lactams such as clavulanate or meropenem resulted in destabilization of PonA1. Our antibiotic profiling experiments indicate that the transpeptidase activity of PonA1 in both M. tuberculosis and M. smegmatis mediates tolerance to specific cell wall-targeting antibiotics, particularly to penicillin V and meropenem. Because M. tuberculosis is an important human pathogen, these structural data provide a template to design novel transpeptidase inhibitors to treat tuberculosis infections. DATABASE: Structural data are available in the PDB database under the accession numbers 5CRF and 5CXW. SN - 1742-4658 UR - https://www.unboundmedicine.com/medline/citation/27101811/Crystal_structures_of_the_transpeptidase_domain_of_the_Mycobacterium_tuberculosis_penicillin-binding_protein_PonA1_reveal_potential_mechanisms_of_antibiotic_resistance. DB - PRIME DP - Unbound Medicine ER -