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A synthetic peptide derived from the non-structural protein 3 of hepatitis C virus serves as a specific substrate for PKC

Standard

A synthetic peptide derived from the non-structural protein 3 of hepatitis C virus serves as a specific substrate for PKC. / Borowski, P; Resch, K; Schmitz, H; Heiland, M.

in: BIOL CHEM, Jahrgang 381, Nr. 1, 01.01.2000, S. 19-27.

Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung

Harvard

Borowski, P, Resch, K, Schmitz, H & Heiland, M 2000, 'A synthetic peptide derived from the non-structural protein 3 of hepatitis C virus serves as a specific substrate for PKC', BIOL CHEM, Jg. 381, Nr. 1, S. 19-27. https://doi.org/10.1515/BC.2000.003

APA

Borowski, P., Resch, K., Schmitz, H., & Heiland, M. (2000). A synthetic peptide derived from the non-structural protein 3 of hepatitis C virus serves as a specific substrate for PKC. BIOL CHEM, 381(1), 19-27. https://doi.org/10.1515/BC.2000.003

Vancouver

Borowski P, Resch K, Schmitz H, Heiland M. A synthetic peptide derived from the non-structural protein 3 of hepatitis C virus serves as a specific substrate for PKC. BIOL CHEM. 2000 Jan 1;381(1):19-27. https://doi.org/10.1515/BC.2000.003

Bibtex

@article{803e54c219224f3692479bbc337ef8ab,
title = "A synthetic peptide derived from the non-structural protein 3 of hepatitis C virus serves as a specific substrate for PKC",
abstract = "A synthetic peptide corresponding to the amino acid sequence Arg1487-Arg-Gly-Arg-Thr-Gly-Arg-Gly-Arg-Arg-Gly-Ile-Tyr-Arg1500 of the hepatitis C virus (HCV) polyprotein was found to be a selective substrate for protein kinase C (PKC). In the presence of Ca2+, TPA and phospholipid, PKC phosphorylates the peptide [termed HCV(1487-1500)] with a Km of 11 microM and Vmax of 24 micromol x min(-1) x mg(-1). HCV(1487-1500) acts as a competitive inhibitor of PKC towards other peptide or protein substrates and inhibits the kinase activity with an IC50 corresponding to the Km values measured for the substrates. N- or C-terminally deleted analogs of HCV(1487-1500) did not show inhibitory effects and were only marginally or not phosphorylatable. We designed an additional peptide in which the tyrosine residue was replaced by phenylalanine ([Phe1499]HCV(1487-1500)). This peptide was neither phosphorylated by other serine/threonine kinases tested nor by whole cell extracts prepared from PKC-depleted cells. [Phe1499]HCV(1487-1500) was used to monitor the TPA-induced translocation of PKC activity to the particulate fraction in JB6 cells. The use of SDS/PAGE to separate the peptide from ATP and Pi allowed to monitor simultaneously PKC autophosphorylation and phosphorylation of the peptide. The data presented here show that[Phe1499]HCV(1487-1500) can serve as a convenient tool for investigations of PKC activity also in the presence of other kinases in tissues or in crude cell extracts.",
keywords = "Amino Acid Sequence, Animals, Binding, Competitive, Cell Line, Cell Membrane, Cytosol, Mice, Peptide Fragments, Phosphorylation, Protein Kinase C, Rats, Substrate Specificity, Tetradecanoylphorbol Acetate, Viral Nonstructural Proteins",
author = "P Borowski and K Resch and H Schmitz and M Heiland",
year = "2000",
month = jan,
day = "1",
doi = "10.1515/BC.2000.003",
language = "English",
volume = "381",
pages = "19--27",
journal = "BIOL CHEM",
issn = "1431-6730",
publisher = "Walter de Gruyter GmbH & Co. KG",
number = "1",

}

RIS

TY - JOUR

T1 - A synthetic peptide derived from the non-structural protein 3 of hepatitis C virus serves as a specific substrate for PKC

AU - Borowski, P

AU - Resch, K

AU - Schmitz, H

AU - Heiland, M

PY - 2000/1/1

Y1 - 2000/1/1

N2 - A synthetic peptide corresponding to the amino acid sequence Arg1487-Arg-Gly-Arg-Thr-Gly-Arg-Gly-Arg-Arg-Gly-Ile-Tyr-Arg1500 of the hepatitis C virus (HCV) polyprotein was found to be a selective substrate for protein kinase C (PKC). In the presence of Ca2+, TPA and phospholipid, PKC phosphorylates the peptide [termed HCV(1487-1500)] with a Km of 11 microM and Vmax of 24 micromol x min(-1) x mg(-1). HCV(1487-1500) acts as a competitive inhibitor of PKC towards other peptide or protein substrates and inhibits the kinase activity with an IC50 corresponding to the Km values measured for the substrates. N- or C-terminally deleted analogs of HCV(1487-1500) did not show inhibitory effects and were only marginally or not phosphorylatable. We designed an additional peptide in which the tyrosine residue was replaced by phenylalanine ([Phe1499]HCV(1487-1500)). This peptide was neither phosphorylated by other serine/threonine kinases tested nor by whole cell extracts prepared from PKC-depleted cells. [Phe1499]HCV(1487-1500) was used to monitor the TPA-induced translocation of PKC activity to the particulate fraction in JB6 cells. The use of SDS/PAGE to separate the peptide from ATP and Pi allowed to monitor simultaneously PKC autophosphorylation and phosphorylation of the peptide. The data presented here show that[Phe1499]HCV(1487-1500) can serve as a convenient tool for investigations of PKC activity also in the presence of other kinases in tissues or in crude cell extracts.

AB - A synthetic peptide corresponding to the amino acid sequence Arg1487-Arg-Gly-Arg-Thr-Gly-Arg-Gly-Arg-Arg-Gly-Ile-Tyr-Arg1500 of the hepatitis C virus (HCV) polyprotein was found to be a selective substrate for protein kinase C (PKC). In the presence of Ca2+, TPA and phospholipid, PKC phosphorylates the peptide [termed HCV(1487-1500)] with a Km of 11 microM and Vmax of 24 micromol x min(-1) x mg(-1). HCV(1487-1500) acts as a competitive inhibitor of PKC towards other peptide or protein substrates and inhibits the kinase activity with an IC50 corresponding to the Km values measured for the substrates. N- or C-terminally deleted analogs of HCV(1487-1500) did not show inhibitory effects and were only marginally or not phosphorylatable. We designed an additional peptide in which the tyrosine residue was replaced by phenylalanine ([Phe1499]HCV(1487-1500)). This peptide was neither phosphorylated by other serine/threonine kinases tested nor by whole cell extracts prepared from PKC-depleted cells. [Phe1499]HCV(1487-1500) was used to monitor the TPA-induced translocation of PKC activity to the particulate fraction in JB6 cells. The use of SDS/PAGE to separate the peptide from ATP and Pi allowed to monitor simultaneously PKC autophosphorylation and phosphorylation of the peptide. The data presented here show that[Phe1499]HCV(1487-1500) can serve as a convenient tool for investigations of PKC activity also in the presence of other kinases in tissues or in crude cell extracts.

KW - Amino Acid Sequence

KW - Animals

KW - Binding, Competitive

KW - Cell Line

KW - Cell Membrane

KW - Cytosol

KW - Mice

KW - Peptide Fragments

KW - Phosphorylation

KW - Protein Kinase C

KW - Rats

KW - Substrate Specificity

KW - Tetradecanoylphorbol Acetate

KW - Viral Nonstructural Proteins

U2 - 10.1515/BC.2000.003

DO - 10.1515/BC.2000.003

M3 - SCORING: Journal article

C2 - 10722046

VL - 381

SP - 19

EP - 27

JO - BIOL CHEM

JF - BIOL CHEM

SN - 1431-6730

IS - 1

ER -