Phosphorylation of the complement component, C9, by an ecto-protein kinase of human leukemic cells

Yoav Paas, Osnat Bohana-Kashtan, Zvi Fishelson

Research output: Contribution to journalArticlepeer-review

Abstract

Ecto-protein kinases (ecto-PK) are surface constituents of many, if not all, animal cell types; normal, transformed or malignant. The occurrence of ecto-PK on the surface of human leukemia cell lines was described. These ecto-PKs have been shown to phosphorylate several exogenous substrates, including the complement C9 protein, an essential component of the terminal complement system. C9 is phosphorylated by ecto-PK of K562 cells on serine residue(s). Phosphorylation occurs in the N-terminal C9a portion produced by cleavage of phosphorylated C9 with human α-thrombin. C9 polymers generated upon incubation of C9 with ZnCl2 do not serve as substrates for the K562 ecto-PK. In contrast, unfolded C9, obtained by reduction and alkylation, serves as a superior substrate for the K562 ecto-PK. Native C9 phosphorylation produced a rather low stoichiometry of incorporated phosphate (around 3%) per C9. Despite that, the phosphorylated C9 expressed reduced hemolytic activity. The complement-sensitive variant of K562 (K562/S) did not express the C9 phosphorylating activity. Various PK inhibitors tested failed to block C9 phosphorylation. Only heparin and 2,3-diphosphoglycerate (dpGA) prevented C9 phosphorylation, indicating that the ecto-PK is related to the casein kinase CK2. C9 can be phosphorylated by ecto-PK from other tumor cells, including Jurkat, SK-OV-3 and BT-474. These results suggest that extracellular phosphorylation of C9 may serve as a protective mechanism against complement in tumor cells.

Original languageEnglish
Pages (from-to)175-185
Number of pages11
JournalImmunopharmacology
Volume42
Issue number1-3
DOIs
StatePublished - May 1999

Keywords

  • C9
  • CK2
  • Complement
  • Ecto-PK
  • Phosphorylation

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