Original scientific paper

Arab-1, a GDSL Lipase from the Model Plant, Arabidopsis thaliana (L.) Heynh.

Goran Mikleušević ; Department of Physical Chemistry, Ruđer Bošković Institute, P. O. Box 180, 10002 Zagreb, Croatia
Branka Salopek-Sondi ; Department of Molecular Biology, Ruđer Bošković Institute, P. O. Box 180, 10002 Zagreb, Croatia
Marija Luić ; Department of Physical Chemistry, Ruđer Bošković Institute, P. O. Box 180, 10002 Zagreb, Croatia

Abstract

Bioinformatics analysis of the protein Arab-1 from model plant Arabidopsis thaliana (L.) Heynh. was performed at the level of primary, secondary and tertiary structure. Results suggest that Arab-1 belongs to the important but structurally very poorely characterised GDSL (SGNH) family of hydrolases. The amino acids Ser42, Gly118, Asn180, Asp346 and His349 are predicted as the active center residues. According to phylogenetic analysis, Arab-1 is related to Brassica napus lipases. Attempts to produce recombinant Arab-1, using pET and pRSET expression systems, and several strains of Escherichia coli cells are described; the procedures were found to be inadequate for the production of amounts of pro-tein sufficient for crystallisation. The problems observed in protein production are discussed in view of bioinformatics analyses and calculated physico-chemical parameters.

Keywords

SGNH (GDSL) hydrolase; lipase; Arabidopsis thaliana; heterologous protein expression; bioinformatics analysis; phylogenetic analysis

Hrčak ID:

39661

URI

https://hrcak.srce.hr/39661

Publication date:

15.7.2009.

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