Abstract
N(G)-Methylation of arginine residues in many nucleic-acid-binding proteins are formed post-translationally, catalysed by S-adenosylmethionine: protein-arginine N-methyltransferase in their glycine-rich and arginine-rich motifs. The amino acid sequences of the stimulator of HIV-1 TAR (Tat-responsive element) RNA-binding protein (SRB) and fibronectin also show the presence of the internal -Gly-Arg-Gly- (-GRG-) sequence, which is potentially methylatable by the methyltransferase. To investigate the sequence requirement for methylation of these proteins, several synthetic oligopeptides with different chain lengths and sequences similar to the -GRG- regions of SRB and fibronectin were synthesized. Whereas the heptapeptide AGGRGKG (residues 16-22 in SRB) served as the methyl acceptor for the methyltransferase with a K(m) of 50 μM, the 19 mer peptide (residues 10-28 in SRB) was methylated with a K(m) of 8.3 μM, indicating that a greater peptide chain length yields a better methyl acceptor. Product analysis of the methylated [methyl-14C]SRB-peptide by HPLC indicated the formation of N(G)-monomethylarginine and N(G),N(G)-dimethyl(asymmetric)- arginine. Synthetic peptides containing the cell attachment sequence [Arg-Gly-Asp ('RGD')] in fibronectin, GRGDSPK, GGRGDSPK and GGGRGDSPK, were also studied; whereas GRGDSPK was a poor methyl acceptor, the longer peptides were better methyl accepters. To provide an understanding of the effect of methylation on fibronectin peptide, arginine-unmethylated and methylated GGRGDSPK were compared for their effect on the mitogenesis induced by β-hexosaminidase A and an agonistic antibody (mAb15) in bovine tracheal smooth-muscle cells; whereas the former inhibited 35-67% of mitogenesis at a concentration of 5-10 μM, the latter did not block mitogenesis. This lack of inhibition by the insertion of a methyl group on the arginyl residue of the cell attachment sequence might be due to the hindrance of the binding of fibronectin peptide to integrins.
| Original language | English |
|---|---|
| Pages (from-to) | 573-578 |
| Number of pages | 6 |
| Journal | Biochemical Journal |
| Volume | 348 |
| Issue number | 3 |
| DOIs | |
| Publication status | Published - 2000 Jun 15 |
Fingerprint
Keywords
- Fibronectin-peptide
- N(G)-methylarginine
- Protein methylase I
- RNA-binding protein
- S-adenosylmethionine
ASJC Scopus subject areas
- Biochemistry
Cite this
Enzymic methylation of arginyl residues in -Gly-Arg-Gly- peptides. / Hyun, Young Lan; Lew, D. Betty; Park, Seung Hee; Kim, Chan Wha; Paik, Woon Ki; Kim, Sangduk.
In: Biochemical Journal, Vol. 348, No. 3, 15.06.2000, p. 573-578.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Enzymic methylation of arginyl residues in -Gly-Arg-Gly- peptides
AU - Hyun, Young Lan
AU - Lew, D. Betty
AU - Park, Seung Hee
AU - Kim, Chan Wha
AU - Paik, Woon Ki
AU - Kim, Sangduk
PY - 2000/6/15
Y1 - 2000/6/15
N2 - N(G)-Methylation of arginine residues in many nucleic-acid-binding proteins are formed post-translationally, catalysed by S-adenosylmethionine: protein-arginine N-methyltransferase in their glycine-rich and arginine-rich motifs. The amino acid sequences of the stimulator of HIV-1 TAR (Tat-responsive element) RNA-binding protein (SRB) and fibronectin also show the presence of the internal -Gly-Arg-Gly- (-GRG-) sequence, which is potentially methylatable by the methyltransferase. To investigate the sequence requirement for methylation of these proteins, several synthetic oligopeptides with different chain lengths and sequences similar to the -GRG- regions of SRB and fibronectin were synthesized. Whereas the heptapeptide AGGRGKG (residues 16-22 in SRB) served as the methyl acceptor for the methyltransferase with a K(m) of 50 μM, the 19 mer peptide (residues 10-28 in SRB) was methylated with a K(m) of 8.3 μM, indicating that a greater peptide chain length yields a better methyl acceptor. Product analysis of the methylated [methyl-14C]SRB-peptide by HPLC indicated the formation of N(G)-monomethylarginine and N(G),N(G)-dimethyl(asymmetric)- arginine. Synthetic peptides containing the cell attachment sequence [Arg-Gly-Asp ('RGD')] in fibronectin, GRGDSPK, GGRGDSPK and GGGRGDSPK, were also studied; whereas GRGDSPK was a poor methyl acceptor, the longer peptides were better methyl accepters. To provide an understanding of the effect of methylation on fibronectin peptide, arginine-unmethylated and methylated GGRGDSPK were compared for their effect on the mitogenesis induced by β-hexosaminidase A and an agonistic antibody (mAb15) in bovine tracheal smooth-muscle cells; whereas the former inhibited 35-67% of mitogenesis at a concentration of 5-10 μM, the latter did not block mitogenesis. This lack of inhibition by the insertion of a methyl group on the arginyl residue of the cell attachment sequence might be due to the hindrance of the binding of fibronectin peptide to integrins.
AB - N(G)-Methylation of arginine residues in many nucleic-acid-binding proteins are formed post-translationally, catalysed by S-adenosylmethionine: protein-arginine N-methyltransferase in their glycine-rich and arginine-rich motifs. The amino acid sequences of the stimulator of HIV-1 TAR (Tat-responsive element) RNA-binding protein (SRB) and fibronectin also show the presence of the internal -Gly-Arg-Gly- (-GRG-) sequence, which is potentially methylatable by the methyltransferase. To investigate the sequence requirement for methylation of these proteins, several synthetic oligopeptides with different chain lengths and sequences similar to the -GRG- regions of SRB and fibronectin were synthesized. Whereas the heptapeptide AGGRGKG (residues 16-22 in SRB) served as the methyl acceptor for the methyltransferase with a K(m) of 50 μM, the 19 mer peptide (residues 10-28 in SRB) was methylated with a K(m) of 8.3 μM, indicating that a greater peptide chain length yields a better methyl acceptor. Product analysis of the methylated [methyl-14C]SRB-peptide by HPLC indicated the formation of N(G)-monomethylarginine and N(G),N(G)-dimethyl(asymmetric)- arginine. Synthetic peptides containing the cell attachment sequence [Arg-Gly-Asp ('RGD')] in fibronectin, GRGDSPK, GGRGDSPK and GGGRGDSPK, were also studied; whereas GRGDSPK was a poor methyl acceptor, the longer peptides were better methyl accepters. To provide an understanding of the effect of methylation on fibronectin peptide, arginine-unmethylated and methylated GGRGDSPK were compared for their effect on the mitogenesis induced by β-hexosaminidase A and an agonistic antibody (mAb15) in bovine tracheal smooth-muscle cells; whereas the former inhibited 35-67% of mitogenesis at a concentration of 5-10 μM, the latter did not block mitogenesis. This lack of inhibition by the insertion of a methyl group on the arginyl residue of the cell attachment sequence might be due to the hindrance of the binding of fibronectin peptide to integrins.
KW - Fibronectin-peptide
KW - N(G)-methylarginine
KW - Protein methylase I
KW - RNA-binding protein
KW - S-adenosylmethionine
UR - http://www.scopus.com/inward/record.url?scp=0034659805&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0034659805&partnerID=8YFLogxK
U2 - 10.1042/0264-6021:3480573
DO - 10.1042/0264-6021:3480573
M3 - Article
VL - 348
SP - 573
EP - 578
JO - Biochemical Journal
JF - Biochemical Journal
SN - 0264-6021
IS - 3
ER -