Crystallization Trials Of Globin Domain Of Environmental Stress Protein Rsbr From Saprospira Grandis
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Date
2011-02
Authors
Badai, Siti Suriawati
Journal Title
Journal ISSN
Volume Title
Publisher
Universiti Sains Malaysia
Abstract
RsbR from the coastal scavenger bacterium Saprospira grandis consists of
two structural domains, the N-terminal globin domain (residues 12-157) and Cterminal
STAS domain (residues 165-276). Observation of the stressosome structure
indicates that the globin fold of the N-terminal domain of RsbR might play crucial
role as stress sensor in the environmental stress signalling. In this study, we have
cloned the gene fragment encoding the globin domain of RsbR and expressed,
purified, and crystallized the recombinant protein. Initial crystallization screening of
the globin domain of RsbR yielded round shaped crystals. The poor morphology of
this crystal was further optimized by screening the pH, analyzing the effect of buffer
choice on crystal quality, testing a reasonable range of precipitant concentration,
trying different salts from the same chemical class, setting crystallization screens at
different temperatures, streak seeding, and screening with the Detergent Screen HT
kit. However, no improvement on crystal quality was observed. Four additional
recombinant variants of N-RsbR domain were constructed and crystallization trials
are currently being performed.
Description
Keywords
Globin Domain Of Environmental Stress Protein Rsbr , Saprospira Grandis