Abstract
Prenylated protein methyltransferase, an enzyme involved in the post-translational modification of many signalling proteins, has been characterized in a parasitic flagellated protozoan, Leishmania donovani. The activity of this enzyme was monitored by the methylation of an artificial substrate, an S-prenylated cysteine analogue, with S-adenosyl-L-[methyl-3H]methionine as methyl donor. More than 85% of the methyltransferase activity was associated with membranes. The enzyme methylates N-acetyl-S-trans, trans-farnesyl-L-cysteine and N-acetyl-S-all-trans-geranylgeranyl-L-cysteine, but N-acetyl-S-trans, trans-geranyl-L-cysteine only very weakly. In contrast with the enzyme from mammals, the leishmanial enzyme had a greater affinity for the farnesylated substrate than for the geranylgeranylated one. Activity in vitro was not modulated by cAMP, protein kinase C activator or guanosine 5'-[γ-thio]triphosphate. An analysis of the endo gene us substrates showed that the carboxymethylated proteins were also isoprenylated. The main carboxymethylated proteins have molecular masses of 95, 68, 55, 46, 34-23, 18 and less than 14 kDa. Treatment of cells with N-acetyl-S-trans, trans-farnesyl-L-cysteine decreased the carboxymethylation level, whereas treatment with guanosine 5'-[γ-thio]triphosphate increased the carboxymethylation of various proteins, particularly those of molecular masses 30-20 kDa.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 513-518 |
| Number of pages | 6 |
| Journal | Biochemical Journal |
| Volume | 342 |
| Issue number | 3 |
| DOIs | |
| State | Published - Sep 15 1999 |
| Externally published | Yes |
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Keywords
- Kinetoplastidae
- Leishmania donovani
- Promastigote
- Protozoans
- Sinefungin
ASJC Scopus subject areas
- Biochemistry
Cite this
Characterization of prenylated protein methyltransferase in Leishmania. / Pierre Hasne, Marie; Lawrence, Françoise.
In: Biochemical Journal, Vol. 342, No. 3, 15.09.1999, p. 513-518.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Characterization of prenylated protein methyltransferase in Leishmania
AU - Pierre Hasne, Marie
AU - Lawrence, Françoise
PY - 1999/9/15
Y1 - 1999/9/15
N2 - Prenylated protein methyltransferase, an enzyme involved in the post-translational modification of many signalling proteins, has been characterized in a parasitic flagellated protozoan, Leishmania donovani. The activity of this enzyme was monitored by the methylation of an artificial substrate, an S-prenylated cysteine analogue, with S-adenosyl-L-[methyl-3H]methionine as methyl donor. More than 85% of the methyltransferase activity was associated with membranes. The enzyme methylates N-acetyl-S-trans, trans-farnesyl-L-cysteine and N-acetyl-S-all-trans-geranylgeranyl-L-cysteine, but N-acetyl-S-trans, trans-geranyl-L-cysteine only very weakly. In contrast with the enzyme from mammals, the leishmanial enzyme had a greater affinity for the farnesylated substrate than for the geranylgeranylated one. Activity in vitro was not modulated by cAMP, protein kinase C activator or guanosine 5'-[γ-thio]triphosphate. An analysis of the endo gene us substrates showed that the carboxymethylated proteins were also isoprenylated. The main carboxymethylated proteins have molecular masses of 95, 68, 55, 46, 34-23, 18 and less than 14 kDa. Treatment of cells with N-acetyl-S-trans, trans-farnesyl-L-cysteine decreased the carboxymethylation level, whereas treatment with guanosine 5'-[γ-thio]triphosphate increased the carboxymethylation of various proteins, particularly those of molecular masses 30-20 kDa.
AB - Prenylated protein methyltransferase, an enzyme involved in the post-translational modification of many signalling proteins, has been characterized in a parasitic flagellated protozoan, Leishmania donovani. The activity of this enzyme was monitored by the methylation of an artificial substrate, an S-prenylated cysteine analogue, with S-adenosyl-L-[methyl-3H]methionine as methyl donor. More than 85% of the methyltransferase activity was associated with membranes. The enzyme methylates N-acetyl-S-trans, trans-farnesyl-L-cysteine and N-acetyl-S-all-trans-geranylgeranyl-L-cysteine, but N-acetyl-S-trans, trans-geranyl-L-cysteine only very weakly. In contrast with the enzyme from mammals, the leishmanial enzyme had a greater affinity for the farnesylated substrate than for the geranylgeranylated one. Activity in vitro was not modulated by cAMP, protein kinase C activator or guanosine 5'-[γ-thio]triphosphate. An analysis of the endo gene us substrates showed that the carboxymethylated proteins were also isoprenylated. The main carboxymethylated proteins have molecular masses of 95, 68, 55, 46, 34-23, 18 and less than 14 kDa. Treatment of cells with N-acetyl-S-trans, trans-farnesyl-L-cysteine decreased the carboxymethylation level, whereas treatment with guanosine 5'-[γ-thio]triphosphate increased the carboxymethylation of various proteins, particularly those of molecular masses 30-20 kDa.
KW - Kinetoplastidae
KW - Leishmania donovani
KW - Promastigote
KW - Protozoans
KW - Sinefungin
UR - http://www.scopus.com/inward/record.url?scp=0033568609&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0033568609&partnerID=8YFLogxK
U2 - 10.1042/0264-6021:3420513
DO - 10.1042/0264-6021:3420513
M3 - Article
C2 - 10477261
AN - SCOPUS:0033568609
VL - 342
SP - 513
EP - 518
JO - Biochemical Journal
JF - Biochemical Journal
SN - 0264-6021
IS - 3
ER -