Isoform-dependent feedback regulation of serine O-acetyltransferase isoenzymes involved in L-cysteine biosynthesis of Entamoeba histolytica.
Serine acetyltransferase (SAT; EC 2.3.1.30) catalyzes the CoA-dependent acetylation of the side chain hydroxyl group of l-serine to form O-acetyl serine, in the first step of the L-cysteine biosynthetic pathway. Since this pathway is selectively present in a few parasitic protists and absent in mammals, it represents a reasonable target to develop new chemotherapeutics. Entamoeba histolytica apparently possesses three SAT isotypes (EhSAT1-3) showing 48-73% mutual identity, a calculated molecular mass of 34.4-37.7 kDa, and an isoelectric point of 5.70-6.63. To better understand the role of individual SAT isotypes, we determined kinetic and inhibitory parameters of recombinant SAT isotypes. While the three SAT isotypes showed comparable Km and k(cat) for L-serine and acetyl-CoA, they showed remarkable differences in their sensitivity to inhibition by L-cysteine. The Ki values for L-cysteine varied by 100-fold (4.7-460 microM) among SAT isotypes (EhSAT1<EhSAT2<EhSAT3). Consequently, these EhSAT isotypes revealed remarkable differences in activity in the presence of physiological L-serine and L-cysteine concentrations. We propose that multiple SAT isotypes with different properties may play complementary roles in the regulation of the cysteine biosynthetic pathway in E. histolytica under different conditions, e.g. during colonization of the intestine and tissue invasion.
Department of Parasitology, Gunma University Graduate School of Medicine, 3-39-22 Showa-machi, Maebashi 371-8511, Japan.
Sarwar
S
Hussain
Vahab
V
Ali
Ghulam
G
Jeelani
Tomoyoshi
T
Nozaki
D000595:Amino Acid Sequence; D000818:Animals; D003545:Cysteine / Q000096:biosynthesis*; D004748:Entamoeba histolytica / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D015800:Protozoan Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D051050:Serine O-Acetyltransferase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity
D007527:Isoenzymes; D015800:Protozoan Proteins; D051050:Serine O-Acetyltransferase; D003545:Cysteine