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Dissecting the Stru...
Dissecting the Structural and Chemical Determinants of the “Open-to-Closed” Motion in the Mannosyltransferase PimA from Mycobacteria
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Rodrigo-Unzueta, Ane (author)
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Ghirardello, Mattia (author)
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Urresti, Saioa (author)
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Delso, Ignacio (author)
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Giganti, David (author)
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Anso, Itxaso (author)
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Trastoy, Beatriz (author)
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Comino, Natalia (author)
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Tersa, Montse (author)
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D'Angelo, Cecilia (author)
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Cifuente, Javier O. (author)
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Marina, Alberto (author)
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- Liebau, Jobst (author)
- Stockholms universitet,Institutionen för biokemi och biofysik
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- Mäler, Lena (author)
- Umeå universitet,Stockholms universitet,Institutionen för biokemi och biofysik,Umeå University, Sweden,Kemiska institutionen
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Chenal, Alexandre (author)
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Albesa-Jové, David (author)
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Merino, Pedro (author)
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Guerin, Marcelo E. (author)
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(creator_code:org_t)
- 2020-07-28
- 2020
- English.
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In: Biochemistry. - : American Chemical Society (ACS). - 0006-2960 .- 1520-4995. ; 59:32, s. 2934-2945
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Abstract
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- The phosphatidyl-myo-inositol mannosyltransferase A (PimA) is an essential peripheral membrane glycosyltransferase that initiates the biosynthetic pathway of phosphatidyl-myo-inositol mannosides (PIMs), key structural elements and virulence factors of Mycobacterium tuberculosis. PimA undergoes functionally important conformational changes, including (i) α-helix-to-β-strand and β-strand-to-α-helix transitions and (ii) an “open-to-closed” motion between the two Rossmann-fold domains, a conformational change that is necessary to generate a catalytically competent active site. In previous work, we established that GDP-Man and GDP stabilize the enzyme and facilitate the switch to a more compact active state. To determine the structural contribution of the mannose ring in such an activation mechanism, we analyzed a series of chemical derivatives, including mannose phosphate (Man-P) and mannose pyrophosphate-ribose (Man-PP-RIB), and additional GDP derivatives, such as pyrophosphate ribose (PP-RIB) and GMP, by the combined use of X-ray crystallography, limited proteolysis, circular dichroism, isothermal titration calorimetry, and small angle X-ray scattering methods. Although the β-phosphate is present, we found that the mannose ring, covalently attached to neither phosphate (Man-P) nor PP-RIB (Man-PP-RIB), does promote the switch to the active compact form of the enzyme. Therefore, the nucleotide moiety of GDP-Man, and not the sugar ring, facilitates the “open-to-closed” motion, with the β-phosphate group providing the high-affinity binding to PimA. Altogether, the experimental data contribute to a better understanding of the structural determinants involved in the “open-to-closed” motion not only observed in PimA but also visualized and/or predicted in other glycosyltransfeases. In addition, the experimental data might prove to be useful for the discovery and/or development of PimA and/or glycosyltransferase inhibitors.
Subject headings
- NATURVETENSKAP -- Biologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences (hsv//eng)
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
Publication and Content Type
- ref (subject category)
- art (subject category)
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- By the author/editor
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Rodrigo-Unzueta, ...
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Ghirardello, Mat ...
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Urresti, Saioa
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Delso, Ignacio
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Giganti, David
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Anso, Itxaso
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show more...
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Trastoy, Beatriz
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Comino, Natalia
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Tersa, Montse
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D'Angelo, Cecili ...
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Cifuente, Javier ...
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Marina, Alberto
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Liebau, Jobst
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Mäler, Lena
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Chenal, Alexandr ...
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Albesa-Jové, Da ...
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Merino, Pedro
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Guerin, Marcelo ...
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show less...
- About the subject
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- NATURAL SCIENCES
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NATURAL SCIENCES
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and Biological Scien ...
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- NATURAL SCIENCES
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NATURAL SCIENCES
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and Biological Scien ...
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and Biochemistry and ...
- Articles in the publication
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Biochemistry
- By the university
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Stockholm University
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Umeå University