Glycerol-3-phosphatase from Trypanosoma brucei brucei
Citation:
Úna McCourt, 'Glycerol-3-phosphatase from Trypanosoma brucei brucei', [thesis], Trinity College (Dublin, Ireland). School of Biochemistry and Immunology, 2008, pp 318Abstract:
Glycerol-3-phosphatase activity was found in both bloodstream and procyclic
form o f trypanosomes with a specific activity of 35 nmol/min/mg of protein and 28
nmol/min/mg of protein respectively. The localisation of the enzyme in bloodstream
form of trypanosomes was identified as cytosolic using cell fractionation techniques
followed by the purification of the enzyme using ion exchange chromatography and
size exclusion chromatography. The purified enzyme had a specific activity of 100
μmol/min/mg protein and had an apparent molecular weight of 32.8 kDa estimated by
Tricine SDS-PAGE. Characterisation studies revealed that the enzyme had a
molecular weight of 33 kDa determined using a gel filtration technique which was in
close agreement with that estimated from SDS-PAGE. The pI of glycerol-3-
phosphatase was estimated by chromatofocusing the enzyme using a pH gradient
during elution and the enzyme eluted at a pH of 5.0, which was consistent with values
reported for other cytosolic enzymes. The activity profile of the enzyme was
determined as a function of H+ concentration and showed a sharp optimum pH at 7.
Glycerol-3-phosphatase required Mg2 for activity and was inhibited SO4 2- as well as
Ba2+, Pb2+ and Ca2+. The enzyme has a Km of 2.7 mM for D,L-glycerol-3-phosphate
and 1.6 mM for L-glycerol-3-phosphate. Approximately equal activity was observed
with D,L-glycerol-3-phosphate, glucose-6-phosphate and fructose-6-phosphate
indicating that the enzyme was not specific for glycerol-3-phosphate.
Author: McCourt, Úna
Advisor:
Voorheis, Howard PaulQualification name:
Doctor of Philosophy (Ph.D.)Publisher:
Trinity College (Dublin, Ireland). School of Biochemistry and ImmunologyNote:
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Full text availableKeywords:
Biochemistry, Ph.D., Ph.D. Trinity College DublinMetadata
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