Localization of cyclic GMP-dependent protein kinase in human mononuclear phagocytes

K. B. Pryzwansky, S. Kidao, T. A. Wyatt, W. Reed, T. M. Lincoln

Research output: Contribution to journalArticlepeer-review

31 Scopus citations

Abstract

The presence and physiological role of cGMP-dependent protein kinase (G-kinase) was investigated in human mononuclear phagocytes. Western blots of monocyte extracts revealed a single polypeptide band that comigrated with purified bovine lung G-kinase. G-kinase was localized by immunofluorescence microscopy in freshly isolated adherent human monocytes, monocyte-derived macrophages cultured from 4 to 14 days, and alveolar macrophages. In monocytes, G-kinase was localized in granules or vesicles in the cytoplasm, at the microtubule organizing center, on filaments, and in the nucleus, In monocyte-derived macrophages, intense staining for G-kinase was found in the vicinity of the Golgi, in vesicles throughout the cytoplasm, and diffusely in the nucleus, Dual-label confocal laser scanning microscopy demonstrated that G-kinase was colocalized with the endoplasmic reticulum, For comparison, G-kinase was localized in alveolar macrophages that were adhered from 3 to 30 min, In these cells, G-kinase was prominent within the organelle-rich area pericortical to the nucleus, However, a well-defined area of intense staining was also observed at the cell periphery at early time points during adherence and spreading. Rhodamine-labeled phalloidin showed that this peripheral area was rich in F-actin, Cytochalasin D, but not nocodazole, inhibited G-kinase targeting to the cell margin. Furthermore, the guanylate cyclase inhibitor LY83583 inhibited alveolar macrophage spreading and staining for G-kinase at the cell periphery. These data suggest that G-kinase may play an important role in cGMP-mediated regulation involved in protein processing and cell motility.

Original languageEnglish (US)
Pages (from-to)670-678
Number of pages9
JournalJournal of Leukocyte Biology
Volume57
Issue number4
DOIs
StatePublished - 1995
Externally publishedYes

Keywords

  • Alveolar macrophage
  • Confocal laser scanning microscopy
  • Cyclic GMP
  • Endoplasmic reticulum
  • F-actin

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Cell Biology

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