Human cystathionine gamma-lyase: developmental and in vitro expression of two isoforms.
Cystathionine gamma-lyase (CGL) is the last enzyme of the trans-sulphuration pathway, which converts methionine into cysteine. To study the possible differences in enzymic activity of the two human cystathionine gamma-lyase isoforms characterized earlier, these were separately expressed in human kidney embryonic 293T cells. Furthermore, developmental changes in the expression of the two mRNA forms as well as the enzymic activity in human liver were studied, as it has been postulated that a change in the relative expression of CGL isoforms causes the postnatal increase in CGL activity. Transfection with the longer isoform increased the CGL activity 1.5-fold, while the activity of the cells transfected with the shorter form did not differ from the basal activity. In human liver samples, CGL activity was only detected in adult tissue (68+/-9 nmol of cysteine/h per mg of protein), whereas activity in fetal, premature and full-term neonatal liver tissue was undetectable. In contrast, strong mRNA expression of both mRNA isoforms was detected from the 19th gestational week onwards and the longer form of CGL appeared to be predominant. The expression of the two mRNA forms varied in parallel. In conclusion, we have shown that only cells overexpressing the longer form of CGL have increased activity, and CGL appears to be regulated at the post-transcriptional level during development.
Levonen
A L
AL
Hospital for Children and Adolescents, University of Helsinki, 00029 Helsinki, Finland. levonen@helsinki.fi
Lapatto
R
R
Saksela
M
M
Raivio
K O
KO
D000328:Adult; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D003542:Cystathionine gamma-Lyase / Q000235:genetics* / Q000378:metabolism; D005333:Fetus; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D007231:Infant, Newborn; D007234:Infant, Premature; D007438:Introns; D007527:Isoenzymes / Q000235:genetics; D007668:Kidney; D008099:Liver / Q000201:enzymology; D012333:RNA, Messenger / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic; D014162:Transfection
D007527:Isoenzymes; D012333:RNA, Messenger; D011994:Recombinant Proteins; D003542:Cystathionine gamma-Lyase