Distant Phe345 mutation compromises the stability and activity of Mycobacterium tuberculosis isocitrate lyase by modulating its structural flexibility

Abstract Isocitrate lyase (ICL), a potential anti-tubercular drug target, catalyzes the first step of the glyoxylate shunt. In the present investigation, we studied the conformational flexibility of MtbICL to better understand its stability and catalytic activity. Our biochemical results showed that...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Harish Shukla, Rohit Shukla, Amit Sonkar, Tripti Pandey, Timir Tripathi
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/b886f7c010434bc289f4df6ea8facbd5
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:b886f7c010434bc289f4df6ea8facbd5
record_format	dspace
spelling	oai:doaj.org-article:b886f7c010434bc289f4df6ea8facbd52021-12-02T11:40:53ZDistant Phe345 mutation compromises the stability and activity of Mycobacterium tuberculosis isocitrate lyase by modulating its structural flexibility10.1038/s41598-017-01235-z2045-2322https://doaj.org/article/b886f7c010434bc289f4df6ea8facbd52017-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-01235-zhttps://doaj.org/toc/2045-2322Abstract Isocitrate lyase (ICL), a potential anti-tubercular drug target, catalyzes the first step of the glyoxylate shunt. In the present investigation, we studied the conformational flexibility of MtbICL to better understand its stability and catalytic activity. Our biochemical results showed that a point mutation at Phe345, which is topologically distant (>10 Å) to the active site signature sequence (189KKCGH193), completely abolishes the activity of the enzyme. In depth computational analyses were carried out for understanding the structural alterations using molecular dynamics, time-dependent secondary structure and principal component analysis. The results showed that the mutated residue increased the structural flexibility and induced conformational changes near the active site (residues 170–210) and in the C-terminal lid region (residues 411–428). Both these regions are involved in the catalytic activity of MtbICL. Upon mutation, the residual mobility of the enzyme increased, resulting in a decrease in the stability, which was confirmed by the lower free energy of stabilization in the mutant enzyme suggesting the destabilization in the structure. Our results have both biological importance and chemical novelty. It reveals internal dynamics of the enzyme structure and also suggests that regions other than the active site should be exploited for targeting MtbICL inhibition and development of novel anti-tuberculosis compounds.Harish ShuklaRohit ShuklaAmit SonkarTripti PandeyTimir TripathiNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Harish Shukla Rohit Shukla Amit Sonkar Tripti Pandey Timir Tripathi Distant Phe345 mutation compromises the stability and activity of Mycobacterium tuberculosis isocitrate lyase by modulating its structural flexibility
description	Abstract Isocitrate lyase (ICL), a potential anti-tubercular drug target, catalyzes the first step of the glyoxylate shunt. In the present investigation, we studied the conformational flexibility of MtbICL to better understand its stability and catalytic activity. Our biochemical results showed that a point mutation at Phe345, which is topologically distant (>10 Å) to the active site signature sequence (189KKCGH193), completely abolishes the activity of the enzyme. In depth computational analyses were carried out for understanding the structural alterations using molecular dynamics, time-dependent secondary structure and principal component analysis. The results showed that the mutated residue increased the structural flexibility and induced conformational changes near the active site (residues 170–210) and in the C-terminal lid region (residues 411–428). Both these regions are involved in the catalytic activity of MtbICL. Upon mutation, the residual mobility of the enzyme increased, resulting in a decrease in the stability, which was confirmed by the lower free energy of stabilization in the mutant enzyme suggesting the destabilization in the structure. Our results have both biological importance and chemical novelty. It reveals internal dynamics of the enzyme structure and also suggests that regions other than the active site should be exploited for targeting MtbICL inhibition and development of novel anti-tuberculosis compounds.
format	article
author	Harish Shukla Rohit Shukla Amit Sonkar Tripti Pandey Timir Tripathi
author_facet	Harish Shukla Rohit Shukla Amit Sonkar Tripti Pandey Timir Tripathi
author_sort	Harish Shukla
title	Distant Phe345 mutation compromises the stability and activity of Mycobacterium tuberculosis isocitrate lyase by modulating its structural flexibility
title_short	Distant Phe345 mutation compromises the stability and activity of Mycobacterium tuberculosis isocitrate lyase by modulating its structural flexibility
title_full	Distant Phe345 mutation compromises the stability and activity of Mycobacterium tuberculosis isocitrate lyase by modulating its structural flexibility
title_fullStr	Distant Phe345 mutation compromises the stability and activity of Mycobacterium tuberculosis isocitrate lyase by modulating its structural flexibility
title_full_unstemmed	Distant Phe345 mutation compromises the stability and activity of Mycobacterium tuberculosis isocitrate lyase by modulating its structural flexibility
title_sort	distant phe345 mutation compromises the stability and activity of mycobacterium tuberculosis isocitrate lyase by modulating its structural flexibility
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/b886f7c010434bc289f4df6ea8facbd5
work_keys_str_mv	AT harishshukla distantphe345mutationcompromisesthestabilityandactivityofmycobacteriumtuberculosisisocitratelyasebymodulatingitsstructuralflexibility AT rohitshukla distantphe345mutationcompromisesthestabilityandactivityofmycobacteriumtuberculosisisocitratelyasebymodulatingitsstructuralflexibility AT amitsonkar distantphe345mutationcompromisesthestabilityandactivityofmycobacteriumtuberculosisisocitratelyasebymodulatingitsstructuralflexibility AT triptipandey distantphe345mutationcompromisesthestabilityandactivityofmycobacteriumtuberculosisisocitratelyasebymodulatingitsstructuralflexibility AT timirtripathi distantphe345mutationcompromisesthestabilityandactivityofmycobacteriumtuberculosisisocitratelyasebymodulatingitsstructuralflexibility
_version_	1718395541108293632

Distant Phe345 mutation compromises the stability and activity of Mycobacterium tuberculosis isocitrate lyase by modulating its structural flexibility

Ejemplares similares