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Titolo:
A DNA ligase from the psychrophile Pseudoalteromonas haloplanktis gives insights into the adaptation of proteins to low temperatures
Autore:
Georlette, D; Jonsson, ZO; Van Petegem, F; Chessa, JP; Van Beeumen, J; Huscher, U; Gerday, C;
Indirizzi:
B6A Univ Liege, Inst Chem, Biochem Lab, B-4000 Sart Tilman, Belgium B6A Univ Liege Sart Tilman Belgium B-4000 b, B-4000 Sart Tilman, Belgium Univ Zurich Irchel, Inst Vet Biochem, CH-8057 Zurich, Switzerland Univ Zurich Irchel Zurich Switzerland CH-8057 H-8057 Zurich, Switzerland Brigham & Womens Hosp, Dept Pathol, Boston, MA 02115 USA Brigham & Womens Hosp Boston MA USA 02115 pt Pathol, Boston, MA 02115 USA Dept Biochem Physiol & Microbiol, Lab Prot Biochem & Prot Engn, Ghent, Belgium Dept Biochem Physiol & Microbiol Ghent Belgium rot Engn, Ghent, Belgium
Titolo Testata:
EUROPEAN JOURNAL OF BIOCHEMISTRY
fascicolo: 12, volume: 267, anno: 2000,
pagine: 3502 - 3512
SICI:
0014-2956(200006)267:12<3502:ADLFTP>2.0.ZU;2-L
Fonte:
ISI
Lingua:
ENG
Soggetto:
MULTIPLE SEQUENCE ALIGNMENT; BLUNT-END LIGATION; NUCLEOTIDE-SEQUENCE; ESCHERICHIA-COLI; PROLINE RESIDUES; ALPHA-AMYLASE; ENCODING GENE; THERMOSTABLE OLIGO-1,6-GLUCOSIDASE; ALTEROMONAS-HALOPLANCTIS; ANTARCTIC BACTERIUM;
Keywords:
NAD(+)-dependent DNA ligase; psychrophile; thermophile; structural comparison; overexpression;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
66
Recensione:
Indirizzi per estratti:
Indirizzo: Gerday, C B6A Univ Liege, Inst Chem, Biochem Lab, B-4000 Sart Tilman, Belgium B6A Univ Liege Sart Tilman Belgium B-4000 Sart Tilman, Belgium
Citazione:
D. Georlette et al., "A DNA ligase from the psychrophile Pseudoalteromonas haloplanktis gives insights into the adaptation of proteins to low temperatures", EUR J BIOCH, 267(12), 2000, pp. 3502-3512

Abstract

The cloning, overexpression and characterization of a cold-adapted DNA ligase from the Antarctic sea water bacterium Pseudoalteromonas haloplanktis are described. Protein sequence analysis revealed that the cold-adapted Ph DNA ligase shows a significant level of sequence similarity to other NAD(+)-dependent DNA ligases and contains several previously described sequence motifs. Also, a decreased level of arginine and proline residues in Ph DNA ligase could be involved in the cold-adaptation strategy. Moreover, 3D modelling of the N-terminal domain of Ph DNA ligase clearly indicates that this domain is destabilized compared with its thermophilic homologue. The recombinant Ph DNA ligase was overexpressed in Escherichia coli and purified to homogeneity. Mass spectroscopy experiments indicated that the purified enzymeis mainly in an adenylated form with a molecular mass of 74 593 Da. Ph DNAligase shows similar overall catalytic properties to other NAD(+)-dependent DNA ligases but is a cold-adapted enzyme as its catalytic efficiency (k(cat)/K-m) at low and moderate temperatures is higher than that of its mesophilic counterpart E. coli DNA ligase. A kinetic comparison of three enzymes adapted to different temperatures (P. haloplanktis, E. coli and Thermus scotoductus DNA ligases) indicated that an increased k(cat) is the most important adaptive parameter for enzymatic activity at low temperatures, whereas a decreased K-m for the nicked DNA substrate seems to allow T. scotoductus DNA ligase to work efficiently at high temperatures. Besides being useful for investigation of the adaptation of enzymes to extreme temperatures, P. haloplanktis DNA ligase, which is very efficient at low temperatures, offersa novel tool for biotechnology.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 26/09/20 alle ore 01:40:49