Archives of Pharmacal Research

, Volume 18, Issue 2, pp 90–99 | Cite as

Potential role of protein kinase C on the differentiation of erythroid progenitor cells

  • Sang Jun Lee
  • In Koo Cho
  • In Hoe Huh
  • Yoon Ki Yom
  • Hyung Soo Ann
Research Articles
Received:
  • 23 Downloads

Abstract

The effect of protein kinase C inhibitors, staurosporine and 1-(5-isoquinolinyl sulfonyl)-2-methyl piperazine(H7) onin vitro differentiation of erythroid progenitor cells which were isolated from spleens of mice infected with the anemia-inducing strain of Friend virus were examined. Erythropoietin-mediated differentiation of erythroid progenitor cells, as determined by the incorporation of59Fe into protoporphyrin, was inhibited by staurosporine and H7 in a concentration-dependent manner. Scatchard analysis of the3H-phorbol-12, 13-dibutyrate binding to erythroid progenitor cells revealed that at the high affinity sites the dissociation constant was 22nM and the maximum number of3H-phorbol-12,13-dibutyrate binding sites per cell was approximately 3.7×105. Cytosolic protein kinase C was isolated from erythroid progenitor cells and then purified by sequential column chromatography. Two isoforms of protein kinase C were found. Photoaffinity labeling of the purified protein kinase C samples with3H-phorbol 12-myristate 13-acetate followed by analysis of SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and autofluorography showed radiolabeled 82-KDa peptides. Radiolabeling of the 82-kDa peptides with3H-phorbol 12-myristate 13-acetate was almost completely blocked by excess unlabeled phorbol 12-myristate 13-acetate. Results of phorbol 12-myristate 13-acetate-promoted phosphorylation with the purified protein kinase C samples showed that the phosphorylation of 82-kDa peptides was increased as the concentration of phorbol 12-myristate 13-acetate was increased from 10−8 M to 10−4M. In light of the findings that erythroid progenitor cells possessed an abundance of protein kinase C and that staurosporine and H7 inhibited erythroid differentiation, it seemed likely that protein kinase C would play a role in the erythroid progenitor cell development.

Key words

Erythropoietin Protein kinase C 3H-phorbol-12, 13-dibutyrate Binding Photoaffinity labeling Autophosphorylation Erythroid Progenitor Cell 
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Copyright information

© The Pharmaceutical Society of Korea 1995

Authors and Affiliations

  • Sang Jun Lee
    • 1
  • In Koo Cho
    • 2
  • In Hoe Huh
    • 2
  • Yoon Ki Yom
    • 3
  • Hyung Soo Ann
    • 4
  1. 1.Cental Reserach LaboratoryChong Kun Dang CoSeoulKorea
  2. 2.College of PharmacyChung Ang UniversitySeoulKorea
  3. 3.College of Natural ScienceSoon Chun Hyang UniversityChoongnamKorea
  4. 4.College of PharmacyDong Duck Women's UniversitySeoulKorea

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