Molecular characterization of Plasmodium falciparum S-adenosylmethionine synthetase.
S-Adenosylmethionine (AdoMet) synthetase (SAMS: EC 2.5.1.6) catalyses the formation of AdoMet from methionine and ATP. We have cloned a gene for Plasmodium falciparum AdoMet synthetase (PfSAMS) (GenBank accession no. AF097923), consisting of 1209 base pairs with no introns. The gene encodes a polypeptide (PfSAMS) of 402 amino acids with a molecular mass of 44844 Da, and has an overall base composition of 67% A+T. PfSAMS is probably a single copy gene, and was mapped to chromosome 9. The PfSAMS protein is highly homologous to all other SAMS, including a conserved motif for the phosphate-binding P-loop, HGGGAFSGKD, and the signature hexapeptide, GAGDQG. All the active-site amino acids for the binding of ADP, P(i) and metal ions are similarly preserved, matching entirely those of human hepatic SAMS and Escherichia coli SAMS. Molecular modelling of PfSAMS guided by the X-ray crystal structure of E. coli SAMS indicates that PfSAMS binds ATP/Mg(2+) in a manner similar to that seen in the E. coli SAMS structure. However, the PfSAMS model shows that it can not form tetramers as does E. coli SAMS, and is probably a dimer instead. There was a differential sensitivity towards the inhibition by cycloleucine between the expressed PfSAMS and the human hepatic SAMS with K(i) values of 17 and 10 mM, respectively. Based on phylogenetic analysis using protein parsimony and neighbour-joining algorithms, the malarial PfSAMS is closely related to SAMS of other protozoans and plants.
Chiang
P K
PK
Walter Reed Army Institute of Research, Washington, DC 20307-5100, USA. peter.chaing@na.amedd.army.mil
Chamberlin
M E
ME
Nicholson
D
D
Soubes
S
S
Su
X
X
Subramanian
G
G
Lanar
D E
DE
Prigge
S T
ST
Scovill
J P
JP
Miller
L H
LH
Chou
J Y
JY
D000595:Amino Acid Sequence; D000818:Animals; D001482:Base Composition; D020134:Catalytic Domain; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D003515:Cycloleucine / Q000494:pharmacology; D018076:DNA, Complementary / Q000235:genetics; D019143:Evolution, Molecular; D018628:Gene Dosage; D017125:Genes, Protozoan; D006801:Humans; D008099:Liver / Q000201:enzymology; D008716:Methionine Adenosyltransferase / Q000037:antagonists & inhibitors / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010802:Phylogeny; D010963:Plasmodium falciparum / Q000201:enzymology*; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D003515:Cycloleucine; D008716:Methionine Adenosyltransferase