Characterization of S-adenosylhomocysteine hydrolase from Cryptosporidium parvum.
The S-adenosylhomocysteine hydrolase from the apicomplexan Cryptosporidium parvum (CpSAHH) has been characterized. CpSAHH is a single-copy, intronless gene of 1479 bp encoding a protein of 493 amino acids with a molecular mass of 55.6 kDa. Reverse transcriptase-polymerase chain reaction analysis confirmed that CpSAHH is expressed both in intracellular stages (in C. parvum-infected HCT-8 cells 24 h after infection) and in sporozoites. CpSAHH was expressed in Escherichia coli TB1 cells as a fusion with maltose-binding protein. The recombinant fusion was cleaved by Factor Xa and the enzymatic activity of both the fusion protein and the purified separated CpSAHH was measured. The enzymatic activity of CpSAHH was inhibited by d-eritadenine, S-DHPA and Ara-A.
Ctrnáctá
Vlasta
V
Department of Tropical Medicine, 1st Faculty of Medicine, Charles University in Prague, Prague, Czech Republic. ctrnacta@yahoo.com
Stejskal
Frantisek
F
Keithly
Janet S
JS
Hrdý
Ivan
I
D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology; D043383:Adenosylhomocysteinase / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D016785:Cryptosporidium parvum / Q000201:enzymology* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D018507:Gene Expression Regulation, Developmental; D007438:Introns; D008969:Molecular Sequence Data; D008970:Molecular Weight; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D016053:RNA, Protozoan / Q000096:biosynthesis / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014740:Vidarabine / Q000494:pharmacology
D012333:RNA, Messenger; D016053:RNA, Protozoan; D011993:Recombinant Fusion Proteins; C100029:eritadenine; C016391:9-(2,3-dihydroxypropyl)adenine; D043383:Adenosylhomocysteinase; D014740:Vidarabine; D000225:Adenine