Abstract
As part of our investigation of fatty acid metabolism in Trypanosoma brucei, we have expressed four acyl-CoA synthetase (TbACS) genes in Esherichia coli. The recombinant proteins, with His-tags on their C-termini, were purified to near homogeneity using nickel-chelate affinity chromatography. Although these enzymes are highly homologous, they have distinct specificities for fatty acid chain length. TbACS1 prefers saturated fatty acids in the range C11:0 to C14:0 and TbACS2 prefers shorter fatty acids, mainly C10:0. TbACS3 and 4, which have 95% sequence identity, have similar specificities, favouring fatty acids between C14:0 and C17:0. In addition, TbACS1, 3 and 4 function well with a variety of unsaturated fatty acids.
Original language | English (US) |
---|---|
Pages (from-to) | 757-761 |
Number of pages | 5 |
Journal | Biochemical Journal |
Volume | 358 |
Issue number | 3 |
DOIs | |
State | Published - Sep 15 2001 |
Externally published | Yes |
Keywords
- ACS signature motif
- Enzyme-coupled assay
- Fatty acid chain length
- Glycosyl phosphatidylinositol myristoylation
- Recombinant acyl-CoA synthetases (ACS)
ASJC Scopus subject areas
- Biochemistry
- Molecular Biology
- Cell Biology