Purification and characterization of some metabolic effects of S-neplanocylmethionine

B. T. Keller, R. S. Clark, Anthony Pegg, R. T. Borchardt

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Our laboratory has previously demonstrated that treatment of mouse L929 cells with 1 μM neplanocin A results in the metabolic formation of S-neplanocylmethionine (Keller, B.T., and R.T. Borchardt, Biochem. Biophys. Res. Commun. 120:131-137 (1984)). The present study described an efficient procedure for the purification of this analog from L cells based on its inherent chemical stability in alkaline conditions. Several metabolic effects of S-neplanocylmethionine are also reported. In L cells, S-neplanocylmethionine was determined to have an apparent half-life of 13 hr compared to 1 hr for S-adenosylmethionine during the initial 2 hr of a cycloleucine block. Analysis of polyamine levels in neplanocin A-treated cells showed a 3.8-fold decrease in putrescine and a 1.7-fold decrease in spermidine by 24 hr, reflecting a decrease in the cell growth rate in response to neplanocin A rather than a direct effect of S-neplanocylmethionine on the cellular S-adenosylmethionine decarboxylase. Consistent with these results are our findings that S-neplanocylmethionine does not significantly inhibit purified rat prostate or Escherichia coli S-adenosylmethionine decarboxylase and that [carboxy-14C]S-neplanocylmethionine exhibits no substrate activity with either enzyme. Purified S-neplanocylmethionine was observed to be a weak inhibitor of both S-adenosylmethionine-dependent protein carboxymethyltransferase and lipid methyltransferase in L cell extracts, having an IC50 value of 205 μM (S-adenosylmethionine = 10 μM). Similar studies with [methyl-3H]S-neplanocylmethionine indicate that the analog has little substrate activity in these two L cell methylation reactions and thus appears to act as a poor competitive inhibitor.

Original languageEnglish (US)
Pages (from-to)364-370
Number of pages7
JournalMolecular pharmacology
Volume28
Issue number4
StatePublished - Dec 1 1985

Fingerprint

Adenosylmethionine Decarboxylase
S-Adenosylmethionine
Protein O-Methyltransferase
Cycloleucine
Putrescine
Spermidine
S-neplanocylmethionine
Polyamines
Methyltransferases
Cell Extracts
Methylation
Inhibitory Concentration 50
Half-Life
Prostate
Escherichia coli
Lipids
Enzymes
Growth
neplanocin A

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Pharmacology

Cite this

Keller, B. T. ; Clark, R. S. ; Pegg, Anthony ; Borchardt, R. T. / Purification and characterization of some metabolic effects of S-neplanocylmethionine. In: Molecular pharmacology. 1985 ; Vol. 28, No. 4. pp. 364-370.
@article{41d3c73728cd4fc1b66ff4b7f41c3a20,
title = "Purification and characterization of some metabolic effects of S-neplanocylmethionine",
abstract = "Our laboratory has previously demonstrated that treatment of mouse L929 cells with 1 μM neplanocin A results in the metabolic formation of S-neplanocylmethionine (Keller, B.T., and R.T. Borchardt, Biochem. Biophys. Res. Commun. 120:131-137 (1984)). The present study described an efficient procedure for the purification of this analog from L cells based on its inherent chemical stability in alkaline conditions. Several metabolic effects of S-neplanocylmethionine are also reported. In L cells, S-neplanocylmethionine was determined to have an apparent half-life of 13 hr compared to 1 hr for S-adenosylmethionine during the initial 2 hr of a cycloleucine block. Analysis of polyamine levels in neplanocin A-treated cells showed a 3.8-fold decrease in putrescine and a 1.7-fold decrease in spermidine by 24 hr, reflecting a decrease in the cell growth rate in response to neplanocin A rather than a direct effect of S-neplanocylmethionine on the cellular S-adenosylmethionine decarboxylase. Consistent with these results are our findings that S-neplanocylmethionine does not significantly inhibit purified rat prostate or Escherichia coli S-adenosylmethionine decarboxylase and that [carboxy-14C]S-neplanocylmethionine exhibits no substrate activity with either enzyme. Purified S-neplanocylmethionine was observed to be a weak inhibitor of both S-adenosylmethionine-dependent protein carboxymethyltransferase and lipid methyltransferase in L cell extracts, having an IC50 value of 205 μM (S-adenosylmethionine = 10 μM). Similar studies with [methyl-3H]S-neplanocylmethionine indicate that the analog has little substrate activity in these two L cell methylation reactions and thus appears to act as a poor competitive inhibitor.",
author = "Keller, {B. T.} and Clark, {R. S.} and Anthony Pegg and Borchardt, {R. T.}",
year = "1985",
month = "12",
day = "1",
language = "English (US)",
volume = "28",
pages = "364--370",
journal = "Molecular Pharmacology",
issn = "0026-895X",
publisher = "American Society for Pharmacology and Experimental Therapeutics",
number = "4",

}

Keller, BT, Clark, RS, Pegg, A & Borchardt, RT 1985, 'Purification and characterization of some metabolic effects of S-neplanocylmethionine', Molecular pharmacology, vol. 28, no. 4, pp. 364-370.

Purification and characterization of some metabolic effects of S-neplanocylmethionine. / Keller, B. T.; Clark, R. S.; Pegg, Anthony; Borchardt, R. T.

In: Molecular pharmacology, Vol. 28, No. 4, 01.12.1985, p. 364-370.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Purification and characterization of some metabolic effects of S-neplanocylmethionine

AU - Keller, B. T.

AU - Clark, R. S.

AU - Pegg, Anthony

AU - Borchardt, R. T.

PY - 1985/12/1

Y1 - 1985/12/1

N2 - Our laboratory has previously demonstrated that treatment of mouse L929 cells with 1 μM neplanocin A results in the metabolic formation of S-neplanocylmethionine (Keller, B.T., and R.T. Borchardt, Biochem. Biophys. Res. Commun. 120:131-137 (1984)). The present study described an efficient procedure for the purification of this analog from L cells based on its inherent chemical stability in alkaline conditions. Several metabolic effects of S-neplanocylmethionine are also reported. In L cells, S-neplanocylmethionine was determined to have an apparent half-life of 13 hr compared to 1 hr for S-adenosylmethionine during the initial 2 hr of a cycloleucine block. Analysis of polyamine levels in neplanocin A-treated cells showed a 3.8-fold decrease in putrescine and a 1.7-fold decrease in spermidine by 24 hr, reflecting a decrease in the cell growth rate in response to neplanocin A rather than a direct effect of S-neplanocylmethionine on the cellular S-adenosylmethionine decarboxylase. Consistent with these results are our findings that S-neplanocylmethionine does not significantly inhibit purified rat prostate or Escherichia coli S-adenosylmethionine decarboxylase and that [carboxy-14C]S-neplanocylmethionine exhibits no substrate activity with either enzyme. Purified S-neplanocylmethionine was observed to be a weak inhibitor of both S-adenosylmethionine-dependent protein carboxymethyltransferase and lipid methyltransferase in L cell extracts, having an IC50 value of 205 μM (S-adenosylmethionine = 10 μM). Similar studies with [methyl-3H]S-neplanocylmethionine indicate that the analog has little substrate activity in these two L cell methylation reactions and thus appears to act as a poor competitive inhibitor.

AB - Our laboratory has previously demonstrated that treatment of mouse L929 cells with 1 μM neplanocin A results in the metabolic formation of S-neplanocylmethionine (Keller, B.T., and R.T. Borchardt, Biochem. Biophys. Res. Commun. 120:131-137 (1984)). The present study described an efficient procedure for the purification of this analog from L cells based on its inherent chemical stability in alkaline conditions. Several metabolic effects of S-neplanocylmethionine are also reported. In L cells, S-neplanocylmethionine was determined to have an apparent half-life of 13 hr compared to 1 hr for S-adenosylmethionine during the initial 2 hr of a cycloleucine block. Analysis of polyamine levels in neplanocin A-treated cells showed a 3.8-fold decrease in putrescine and a 1.7-fold decrease in spermidine by 24 hr, reflecting a decrease in the cell growth rate in response to neplanocin A rather than a direct effect of S-neplanocylmethionine on the cellular S-adenosylmethionine decarboxylase. Consistent with these results are our findings that S-neplanocylmethionine does not significantly inhibit purified rat prostate or Escherichia coli S-adenosylmethionine decarboxylase and that [carboxy-14C]S-neplanocylmethionine exhibits no substrate activity with either enzyme. Purified S-neplanocylmethionine was observed to be a weak inhibitor of both S-adenosylmethionine-dependent protein carboxymethyltransferase and lipid methyltransferase in L cell extracts, having an IC50 value of 205 μM (S-adenosylmethionine = 10 μM). Similar studies with [methyl-3H]S-neplanocylmethionine indicate that the analog has little substrate activity in these two L cell methylation reactions and thus appears to act as a poor competitive inhibitor.

UR - http://www.scopus.com/inward/record.url?scp=0022359593&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0022359593&partnerID=8YFLogxK

M3 - Article

VL - 28

SP - 364

EP - 370

JO - Molecular Pharmacology

JF - Molecular Pharmacology

SN - 0026-895X

IS - 4

ER -