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Crystallization and preliminary crystallographic analysis of cysteine synthase from Entamoeba histolytica.


ABSTRACT: Entamoeba histolytica, the causative agent of human amoebiasis, is essentially anaerobic, requiring a small amount of oxygen for growth. It cannot tolerate the higher concentration of oxygen present in human tissues or blood. However, during tissue invasion it is exposed to a higher level of oxygen, leading to oxygen stress. Cysteine, which is a vital thiol in E. histolytica, plays an essential role in its oxygen-defence mechanisms. The major route of cysteine biosynthesis in this parasite is the condensation of O-acetylserine with sulfide by the de novo cysteine-biosynthetic pathway, which involves cysteine synthase (EhCS) as a key enzyme. In this study, EhCS was cloned, expressed in Escherichia coli and purified by affinity and size-exclusion chromatography. The purified protein was crystallized in space group P4(1) with two molecules per asymmetric unit and a complete data set was collected to a resolution of 1.86 A. A molecular-replacement solution was obtained using the Salmonella typhimurium O-acetylserine sulfhydrylase structure as a probe and had a correlation coefficient of 37.7% and an R factor of 48.8%.

SUBMITTER: Krishna C 

PROVIDER: S-EPMC2335080 | biostudies-literature | 2007 Jun

REPOSITORIES: biostudies-literature

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Crystallization and preliminary crystallographic analysis of cysteine synthase from Entamoeba histolytica.

Krishna Chinthalapudi C   Jain Ruchi R   Kashav Tara T   Wadhwa Dinakar D   Alam Neelima N   Gourinath S S  

Acta crystallographica. Section F, Structural biology and crystallization communications 20070512 Pt 6


Entamoeba histolytica, the causative agent of human amoebiasis, is essentially anaerobic, requiring a small amount of oxygen for growth. It cannot tolerate the higher concentration of oxygen present in human tissues or blood. However, during tissue invasion it is exposed to a higher level of oxygen, leading to oxygen stress. Cysteine, which is a vital thiol in E. histolytica, plays an essential role in its oxygen-defence mechanisms. The major route of cysteine biosynthesis in this parasite is th  ...[more]

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