Compartmentalization of cyclic GMP-dependent protein kinase in formyl-peptide stimulated neutrophils

Katherine B. Pryzwansky, Todd A. Wyatt, Harriette Nichols, Thomas M. Lincoln

Research output: Contribution to journalArticle

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Abstract

The presence and physiologic role of cyclic GMP-dependent protein kinase (G-kinase) in human neutrophils was investigated by Western blot analysis and immunocytochemistry. Small quantities of G-kinase were found in the cytoskeletal-enriched fraction of neutrophil lysates as detected by Western blots using a polyclonal antibody raised against bovine aorta G-kinase. Immunofluorescence microscopy demonstrated in adherent neutrophils that G-kinase was localized diffusely within the cytoplasm, at the microtubule organizing center, and in the euchromatin of the nucleus. Because cyclic GMP is implicated as a modulator of neutrophil chemotaxis, G-kinase localization was investigated in neutrophils activated with N-formyl-methionyl-leucyl-phenylalanine (fMLP). fMLP stimulated transient focal changes in G-kinase localization that coincided with transient changes in cell shape. G-kinase translocated over a period of 5 minutes from diffuse staining of the cytosol to filaments within the uropod of polarized cells (1 minute), to bundles of filaments associated with loss of cell polarity (2.5 minutes), and finally to more intense staining of the nuclear euchromatin (5 minutes). Optical sectioning of neutrophils by confocal laser scanning microscopy confirmed that G-kinase was restricted to specific sub-cellular compartments during cell activation. This transient localization of G-kinase was disrupted by cytoskeletal inhibitors and was augmented by 8-Br-cyclic GMP. These data provide evidence for the first time that G-kinase plays a physiologic role in human neutrophils, and support the concept of compartmentalization of cyclic nucleotides during neutrophil activation.

Original languageEnglish (US)
Pages (from-to)612-618
Number of pages7
JournalBlood
Volume76
Issue number3
StatePublished - Aug 1 1990
Externally publishedYes

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Cyclic GMP-Dependent Protein Kinases
Protein Kinases
Neutrophils
Peptides
Euchromatin
N-Formylmethionine Leucyl-Phenylalanine
Microscopic examination
Western Blotting
Chemical activation
Microtubule-Organizing Center
Staining and Labeling
Cell Polarity
Neutrophil Activation
Cell Shape
Cyclic Nucleotides
Cyclic GMP
Chemotaxis
Fluorescence Microscopy
Confocal Microscopy
Cytosol

ASJC Scopus subject areas

  • Hematology

Cite this

Compartmentalization of cyclic GMP-dependent protein kinase in formyl-peptide stimulated neutrophils. / Pryzwansky, Katherine B.; Wyatt, Todd A.; Nichols, Harriette; Lincoln, Thomas M.

In: Blood, Vol. 76, No. 3, 01.08.1990, p. 612-618.

Research output: Contribution to journalArticle

Pryzwansky, KB, Wyatt, TA, Nichols, H & Lincoln, TM 1990, 'Compartmentalization of cyclic GMP-dependent protein kinase in formyl-peptide stimulated neutrophils', Blood, vol. 76, no. 3, pp. 612-618.
Pryzwansky, Katherine B. ; Wyatt, Todd A. ; Nichols, Harriette ; Lincoln, Thomas M. / Compartmentalization of cyclic GMP-dependent protein kinase in formyl-peptide stimulated neutrophils. In: Blood. 1990 ; Vol. 76, No. 3. pp. 612-618.
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