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The HU protein from Thermotoga maritima: recombinant expression, purification and physicochemical characterization of an extremely hyperthermophilic DNA-binding protein.
J Mol Biol. 1999 Sep 03; 291(5):1135-46.JM

Abstract

The histone-like protein TmHU from the hyperthermophilic eubacterium Thermotoga maritima was cloned, expressed to high levels in Escherichia coli, and purified to homogeneity by heat precipitation and cation exchange chromatography. CD spectroscopical studies with secondary structure analysis as well as comparative modeling demonstrate that the dimeric TmHU has a tertiary structure similar to other homologous HU proteins. The Tm of the protein was determined to be 96 degrees C, and thermal unfolding is nearly completely reversible. Surface plasmon resonance measurements for TmHU show that the protein binds to DNA in a highly cooperative manner, with a KD of 73 nM and a Hill coefficient of 7.6 for a 56 bp DNA fragment. It is demonstrated that TmHU is capable to increase the melting point of a synthetic, double-stranded DNA (poly[d(A-T)]) by 47 degrees C, thus suggesting that DNA stabilization may be a major function of this protein in hyperthermophiles. The significant in vitro protection of double-helical DNA may be useful for biotechnological applications.

Authors+Show Affiliations

Institut für Biotechnologie, Martin-Luther-Universität Halle-Wittenberg, Saale, Germany. dirk.esser@biochemtech.uni-halle.deNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

10518949

Citation

Esser, D, et al. "The HU Protein From Thermotoga Maritima: Recombinant Expression, Purification and Physicochemical Characterization of an Extremely Hyperthermophilic DNA-binding Protein." Journal of Molecular Biology, vol. 291, no. 5, 1999, pp. 1135-46.
Esser D, Rudolph R, Jaenicke R, et al. The HU protein from Thermotoga maritima: recombinant expression, purification and physicochemical characterization of an extremely hyperthermophilic DNA-binding protein. J Mol Biol. 1999;291(5):1135-46.
Esser, D., Rudolph, R., Jaenicke, R., & Böhm, G. (1999). The HU protein from Thermotoga maritima: recombinant expression, purification and physicochemical characterization of an extremely hyperthermophilic DNA-binding protein. Journal of Molecular Biology, 291(5), 1135-46.
Esser D, et al. The HU Protein From Thermotoga Maritima: Recombinant Expression, Purification and Physicochemical Characterization of an Extremely Hyperthermophilic DNA-binding Protein. J Mol Biol. 1999 Sep 3;291(5):1135-46. PubMed PMID: 10518949.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - The HU protein from Thermotoga maritima: recombinant expression, purification and physicochemical characterization of an extremely hyperthermophilic DNA-binding protein. AU - Esser,D, AU - Rudolph,R, AU - Jaenicke,R, AU - Böhm,G, PY - 1999/10/16/pubmed PY - 1999/10/16/medline PY - 1999/10/16/entrez SP - 1135 EP - 46 JF - Journal of molecular biology JO - J Mol Biol VL - 291 IS - 5 N2 - The histone-like protein TmHU from the hyperthermophilic eubacterium Thermotoga maritima was cloned, expressed to high levels in Escherichia coli, and purified to homogeneity by heat precipitation and cation exchange chromatography. CD spectroscopical studies with secondary structure analysis as well as comparative modeling demonstrate that the dimeric TmHU has a tertiary structure similar to other homologous HU proteins. The Tm of the protein was determined to be 96 degrees C, and thermal unfolding is nearly completely reversible. Surface plasmon resonance measurements for TmHU show that the protein binds to DNA in a highly cooperative manner, with a KD of 73 nM and a Hill coefficient of 7.6 for a 56 bp DNA fragment. It is demonstrated that TmHU is capable to increase the melting point of a synthetic, double-stranded DNA (poly[d(A-T)]) by 47 degrees C, thus suggesting that DNA stabilization may be a major function of this protein in hyperthermophiles. The significant in vitro protection of double-helical DNA may be useful for biotechnological applications. SN - 0022-2836 UR - https://www.unboundmedicine.com/medline/citation/10518949/The_HU_protein_from_Thermotoga_maritima:_recombinant_expression_purification_and_physicochemical_characterization_of_an_extremely_hyperthermophilic_DNA_binding_protein_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0022-2836(99)93022-X DB - PRIME DP - Unbound Medicine ER -