102168 20230519145441.0 doi 10.3390/ijms22063231 sideral 124111 ART-2021-124111 eng Neira, J.L. The histidine phosphocarrier kinase/phosphorylase from bacillus subtilis is an oligomer in solution with a high thermal stability 2021 Access copy available to the general public Unrestricted The histidine phosphocarrier protein (HPr) kinase/phosphorylase (HPrK/P) modulates the phosphorylation state of the HPr protein, and it is involved in the use of carbon sources by Gram-positive bacteria. Its X-ray structure, as concluded from crystals of proteins from several species, is a hexamer; however, there are no studies about its conformational stability, and how its structure is modified by the pH. We have embarked on the conformational characterization of HPrK/P of Bacillus subtilis (bsHPrK/P) in solution by using several spectroscopic (namely, fluorescence and circular dichroism (CD)) and biophysical techniques (namely, small-angle X-ray-scattering (SAXS) and dynamic light-scattering (DLS)). bsHPrK/P was mainly a hexamer in solution at pH 7.0, in the presence of phosphate. The protein had a high conformational stability, with an apparent thermal denaturation midpoint of ~70¿ C, at pH 7.0, as monitored by fluorescence and CD. The protein was very pH-sensitive, precipitated between pH 3.5 and 6.5; below pH 3.5, it had a molten-globule-like conformation; and it acquired a native-like structure in a narrow pH range (between pH 7.0 and 8.0). Guanidinium hydrochloride (GdmCl) denaturation occurred through an oligomeric intermediate. On the other hand, urea denaturation occurred as a single transition, in the range of concentrations between 1.8 and 18 µM, as detected by far-UV CD and fluorescence. info:eu-repo/grantAgreement/ES/AEI/CTQ2017-85425-P info:eu-repo/grantAgreement/ES/MCIU-AEI-FEDER/RTI2018-097991-B-I00 info:eu-repo/grantAgreement/ES/MINECO/BIO2016-78020-R info:eu-repo/semantics/openAccess by http://creativecommons.org/licenses/by/3.0/es/ 6.208 2021 BIOCHEMISTRY & MOLECULAR BIOLOGY 69 / 297 = 0.232 2021 Q1 T1 CHEMISTRY, MULTIDISCIPLINARY 50 / 180 = 0.278 2021 Q2 T1 1.176 2021 Computer Science Applications 2021 Q1 Inorganic Chemistry 2021 Q1 Spectroscopy 2021 Q1 Organic Chemistry 2021 Q1 Physical and Theoretical Chemistry 2021 Q1 Molecular Biology 2021 Q1 6.9 2021 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Cámara-Artigas, A. Hernández-Cifre, J.G. Ortore, M.G. 22, 6 (2021), 3231 [23 pp.] Int. j. mol. sci. International Journal of Molecular Sciences 1661-6596 2219816 http://zaguan.unizar.es/record/102168/files/texto_completo.pdf Versión publicada 2692668 http://zaguan.unizar.es/record/102168/files/texto_completo.jpg?subformat=icon icon Versión publicada oai:zaguan.unizar.es:102168 articulos driver 2023-05-18-14:32:04 ARTICLE