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Mutational and structural study of RipA, a key enzyme in Mycobacterium tuberculosis cell division: evidence for the L-to-D inversion of configuration of the catalytic cysteine.

Squeglia F, Ruggiero A, Romano M, Vitagliano L, Berisio R
Acta Crystallogr D Biol Crystallogr 70 2295-300 (2014)
Related entries: 4q4g, 4q4t

Cited: 13 times
EuropePMC logo PMID: 25195744

Abstract

RipA is a key cysteine protease of Mycobacterium tuberculosis as it is responsible for bacterial daughter-cell separation. Although it is an important target for antimicrobial development, its mechanism of action and its interaction pattern with its substrate are hitherto unknown. By combining crystallographic and mutational studies with functional assays and molecular modelling, it is shown that the catalytic activity of the enzyme relies on a Cys-His-Glu triad and the impact of the mutation of each residue of the triad on the structure and function of RipA is analysed. Unexpectedly, the crystallographic analyses reveal that mutation of the glutamic acid to alanine results in inversion of the configuration of the catalytic cysteine. The consequent burial of the catalytic cysteine side chain explains the enzyme inactivation upon mutation. These data point to a novel role of the acidic residue often present in the triad of cysteine proteases as a supervisor of cysteine configuration through preservation of the local structural integrity.

Reviews - 4q4n mentioned but not cited (1)

  1. The Cell Wall Hydrolytic NlpC/P60 Endopeptidases in Mycobacterial Cytokinesis: A Structural Perspective. Squeglia F, Moreira M, Ruggiero A, Berisio R. Cells 8 E609 (2019)

Articles - 4q4n mentioned but not cited (1)

  1. Potential Efficacy of β-Amyrin Targeting Mycobacterial Universal Stress Protein by In Vitro and In Silico Approach. Beg MA, Shivangi, Afzal O, Akhtar MS, Altamimi ASA, Hussain A, Imam MA, Ahmad MN, Chopra S, Athar F. Molecules 27 4581 (2022)


Reviews citing this publication (3)

  1. Chemistry of Peptidoglycan in Mycobacterium tuberculosis Life Cycle: An off-the-wall Balance of Synthesis and Degradation. Squeglia F, Ruggiero A, Berisio R. Chemistry 24 2533-2546 (2018)
  2. A structural overview of mycobacterial adhesins: Key biomarkers for diagnostics and therapeutics. Squeglia F, Ruggiero A, De Simone A, Berisio R. Protein Sci 27 369-380 (2018)
  3. Collagen degradation in tuberculosis pathogenesis: the biochemical consequences of hosting an undesired guest. Squeglia F, Ruggiero A, Berisio R. Biochem J 475 3123-3140 (2018)

Articles citing this publication (8)

  1. Adherence and invasive properties of Corynebacterium diphtheriae strains correlates with the predicted membrane-associated and secreted proteome. Sangal V, Blom J, Sutcliffe IC, von Hunolstein C, Burkovski A, Hoskisson PA. BMC Genomics 16 765 (2015)
  2. Structure and transformation of bacteriophage A511 baseplate and tail upon infection of Listeria cells. Guerrero-Ferreira RC, Hupfeld M, Nazarov S, Taylor NM, Shneider MM, Obbineni JM, Loessner MJ, Ishikawa T, Klumpp J, Leiman PG. EMBO J 38 e99455 (2019)
  3. NlpC/P60 domain-containing proteins of Mycobacterium avium subspecies paratuberculosis that differentially bind and hydrolyze peptidoglycan. Bannantine JP, Lingle CK, Adam PR, Ramyar KX, McWhorter WJ, Stabel JR, Picking WD, Geisbrecht BV. Protein Sci 25 840-851 (2016)
  4. Structure and dynamics of the multi-domain resuscitation promoting factor RpfB from Mycobacterium tuberculosis. Ruggiero A, Squeglia F, Romano M, Vitagliano L, De Simone A, Berisio R. J Biomol Struct Dyn 35 1322-1330 (2017)
  5. Enterococcus NlpC/p60 Peptidoglycan Hydrolase SagA Localizes to Sites of Cell Division and Requires Only a Catalytic Dyad for Protease Activity. Espinosa J, Lin TY, Estrella Y, Kim B, Molina H, Hang HC. Biochemistry 59 4470-4480 (2020)
  6. Enzyme characterization and biological activities of a resuscitation promoting factor from an oil degrading bacterium Rhodococcus erythropolis KB1. Luo D, Chen J, Xie G, Yue L, Wang Y. PeerJ 7 e6951 (2019)
  7. Biochemical Characterizations of the Putative Endolysin Ecd09610 Catalytic Domain from Clostridioides difficile. Sekiya H, Yamaji H, Yoshida A, Matsunami R, Kamitori S, Tamai E. Antibiotics (Basel) 11 1131 (2022)
  8. Regulation of the cell division hydrolase RipC by the FtsEX system in Mycobacterium tuberculosis. Li J, Xu X, Shi J, Hermoso JA, Sham LT, Luo M. Nat Commun 14 7999 (2023)


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