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Steinhauf, Daniel, 2010. How to kill a parasite : a report : an inhibitor study of ribose-5-phosphate isomerase B from Giardia lamblia as a possible drug target . First cycle, G2E. Uppsala: SLU, Dept. of Molecular Biology (until 131231)

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Abstract

The parasite Giardia lamblia is known to cause the disease giardiasis, which is a major threat against the public health, especially in developing countries. A recent study has shown that over 200 million people have giardiasis now, and about 500 000 more people are infected every year. A G. lamblia infection is usually treated with the drugs metronidazole, albendazole, tinidazole and furazolidone. These drugs are in some cases toxic and have severe side effects. Therefore new methods of killing the parasite are of great importance. Ribose-5-phosphate isomerase (Rpi) is an enzyme active in the pentose phosphate pathway, and it catalyses the conversion of ribose 5-phosphate to ribulose 5-phosphate and vice versa. Some enzymes in this class can also act on 6-carbon substrates, interconverting allose 6-phosphate and allulose 6-phosphate. To find which inhibitor works best on the enzyme from G. lamblia (GlRpiB), the enzyme was expressed as a His-tagged construct in E. coli was purified then tested to check its purity. When it was confirmed to be the right protein, the protein was used for crystallization and kinetic assays. Allose-6-phosphate was added before crystallization. Crystals formed in the presence of allose-6-phosphate. The Km, Vmax and kcat were determined and inhibitory tests were performed. It was determined that the Km for ribose-5-phosphate is about 10 mM, and the kcat is around 260sec-1. The Km of for allose-6-phosphate is 0.41 mM, and kcat 0.008sec-1. Among 6 substrate-like compounds tested, only 4PEH (4-phospho-D-erythronohydroxamic acid) showed significant inhibition of GlRpiB.

Main title:How to kill a parasite
Subtitle:a report : an inhibitor study of ribose-5-phosphate isomerase B from Giardia lamblia as a possible drug target
Authors:Steinhauf, Daniel
Supervisor:Mowbray, Sherry and Stern, Ana Laura
Examiner:Ståhlberg, Jerry
Series:UNSPECIFIED
Volume/Sequential designation:UNSPECIFIED
Year of Publication:2010
Level and depth descriptor:First cycle, G2E
Student's programme affiliation:NK002 Biotechnology - Bachelor's Programme 180 HEC
Department:(NL, NJ) > Dept. of Molecular Biology (until 131231)
Keywords:Giardia lamblia, Ribose-5-phosphate isomerase, Giardiasis, Inhibitor, Parasite, Diarrhea, Pentose phosphate pathway, 4PEH
URN:NBN:urn:nbn:se:slu:epsilon-8-565
Permanent URL:
http://urn.kb.se/resolve?urn=urn:nbn:se:slu:epsilon-8-565
Subjects:Human medicine, health, and safety
Language:English
Deposited On:26 Jul 2010 12:47
Metadata Last Modified:20 Apr 2012 14:14

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