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Functional studies of rat galactokinase

Paper ID Volume ID Publish Year Pages File Format Full-Text
24793 43537 2009 5 PDF Available
Title
Functional studies of rat galactokinase
Abstract

Galactokinase is an ATP-dependent enzyme that catalyzes the phosphorylation of galactose to form galactose-1-phosphate. The defect in human galactokinase can result in the disease of galactosemia. On the other hand, the control of galactose-1-phosphate production by inhibiting galactokinase is a potential therapy for another disease referred to as classic galactosemia. Many pharmaceutically important compounds derive from carbohydrate-containing natural products, and glycorandomization is one of the most efficient approaches for complex secondary metabolites. Therefore, it is important to further understand the interaction between galactokinase and its substrate or substrate analogs. In the present study, we cloned and purified both N- and C-terminal His-tagged rat galactokinase. We then constructed and purified a variety of variant enzymes, which were studied using kinetics with galactose and its analogs as substrates. We found that the binding of the ATP may induce conformational change to the enzyme so that the enzyme can bind galactose specifically. Asp186 was found to be a possible catalytic residue. The mutants were incubated with fluorescent trinitrophenyl-ATP for the characterization of their ATP binding sites. Various other substrate analogs, aminoglycosides and flavanoids were also tested and found to be competitive inhibitors of rat galactokinase.

Keywords
ATP, adenosine 5′-triphosphate; DMSO, dimethyl sulfoxide; Fmax, maximum fluorescence intensity; GALK, galactokinase; gal-1-p, galactose-1-phosphate; Kd, dissociation constant; NADH, nicotinamide adenine dinucleotide, reduced form; SDS, sodium dodecylsulfa
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Functional studies of rat galactokinase
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 141, Issues 3–4, 20 May 2009, Pages 142–146
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
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Any Questions? feel free to contact us