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NMR characterization of a 264-residue hyperthermostable endo-beta-1,3-glucanase.
Biochem Biophys Res Commun. 2010 Jan 01; 391(1):370-5.BB

Abstract

Insight into the hyperthermostable endo-beta-1,3-glucanase pfLamA from Pyrococcus furiosus is obtained by using NMR spectroscopy. pfLamA functions optimally at 104 degrees C and recently the X-ray structure of pfLamA has been obtained at 20 degrees C, a temperature at which the enzyme is inactive. In this study, near-complete (>99%) NMR assignments are presented of chemical shifts of pfLamA in presence and absence of calcium at 62 degrees C, a temperature at which the enzyme is biologically active. The protein contains calcium and the effects of calcium on the protein are assessed. Calcium binding results in relatively small chemical shift changes in a region distant from the active site of pfLamA and thus causes only minor conformational modifications. Removal of calcium does not significantly alter the denaturation temperature of pfLamA, implying that calcium does not stabilize the enzyme against global unfolding. The data obtained form the basis for elucidation of the molecular origins involved in conformational stability and biological activity of hyperthermophilic endo-beta-1,3-glucanases at extreme temperatures.

Authors+Show Affiliations

Laboratory of Biochemistry, Wageningen University, Dreijenlaan 3, 6703 HA Wageningen, The Netherlands.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

19913513

Citation

Ippel, Johannes H., et al. "NMR Characterization of a 264-residue Hyperthermostable Endo-beta-1,3-glucanase." Biochemical and Biophysical Research Communications, vol. 391, no. 1, 2010, pp. 370-5.
Ippel JH, Koutsopoulos S, Nabuurs SM, et al. NMR characterization of a 264-residue hyperthermostable endo-beta-1,3-glucanase. Biochem Biophys Res Commun. 2010;391(1):370-5.
Ippel, J. H., Koutsopoulos, S., Nabuurs, S. M., van Berkel, W. J., van der Oost, J., & van Mierlo, C. P. (2010). NMR characterization of a 264-residue hyperthermostable endo-beta-1,3-glucanase. Biochemical and Biophysical Research Communications, 391(1), 370-5. https://doi.org/10.1016/j.bbrc.2009.11.065
Ippel JH, et al. NMR Characterization of a 264-residue Hyperthermostable Endo-beta-1,3-glucanase. Biochem Biophys Res Commun. 2010 Jan 1;391(1):370-5. PubMed PMID: 19913513.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - NMR characterization of a 264-residue hyperthermostable endo-beta-1,3-glucanase. AU - Ippel,Johannes H, AU - Koutsopoulos,Sotirios, AU - Nabuurs,Sanne M, AU - van Berkel,Willem J H, AU - van der Oost,John, AU - van Mierlo,Carlo P M, Y1 - 2009/11/12/ PY - 2009/10/27/received PY - 2009/11/07/accepted PY - 2009/11/17/entrez PY - 2009/11/17/pubmed PY - 2010/3/17/medline SP - 370 EP - 5 JF - Biochemical and biophysical research communications JO - Biochem Biophys Res Commun VL - 391 IS - 1 N2 - Insight into the hyperthermostable endo-beta-1,3-glucanase pfLamA from Pyrococcus furiosus is obtained by using NMR spectroscopy. pfLamA functions optimally at 104 degrees C and recently the X-ray structure of pfLamA has been obtained at 20 degrees C, a temperature at which the enzyme is inactive. In this study, near-complete (>99%) NMR assignments are presented of chemical shifts of pfLamA in presence and absence of calcium at 62 degrees C, a temperature at which the enzyme is biologically active. The protein contains calcium and the effects of calcium on the protein are assessed. Calcium binding results in relatively small chemical shift changes in a region distant from the active site of pfLamA and thus causes only minor conformational modifications. Removal of calcium does not significantly alter the denaturation temperature of pfLamA, implying that calcium does not stabilize the enzyme against global unfolding. The data obtained form the basis for elucidation of the molecular origins involved in conformational stability and biological activity of hyperthermophilic endo-beta-1,3-glucanases at extreme temperatures. SN - 1090-2104 UR - https://www.unboundmedicine.com/medline/citation/19913513/NMR_characterization_of_a_264_residue_hyperthermostable_endo_beta_13_glucanase_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0006-291X(09)02240-2 DB - PRIME DP - Unbound Medicine ER -