Esser, Dirk and Rudolph, Rainer and Jaenicke, Rainer and Boehm, Gerald (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). pp. 1135-1146. ISSN 0022-2836,
Full text not available from this repository. (Request a copy)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 T-m 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 K-D of 73 nM and a Kill 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. (C) 1999 Academic Press.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | HISTONE-LIKE PROTEINS; ESCHERICHIA-COLI; BACILLUS-STEAROTHERMOPHILUS; COMPLEX; IHF; THERMOSTABILITY; RECOGNITION; STABILITY; ISOMERASE; CONTAINS; Thermotoga maritima; DNA-binding protein; DNA stability; histone-like protein; hyperthermophilic protein |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biophysik und physikalische Biochemie |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 21 Dec 2022 07:27 |
| Last Modified: | 21 Dec 2022 07:27 |
| URI: | https://pred.uni-regensburg.de/id/eprint/49003 |
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