Characterization of the role of CaMKI-like kinase (CKLiK) in human granulocyte function

• Published in: Blood Vol. 106; no. 3; pp. 1076 - 1083
• Main Authors: Verploegen, Sandra, Ulfman, Laurien, van Deutekom, Hanneke W.M., van Aalst, Corneli, Honing, Henk, Lammers, Jan-Willem J., Koenderman, Leo, Coffer, Paul J.
• Format: Journal Article
• Language: English
• Published: Washington, DC Elsevier Inc 01.08.2005; The Americain Society of Hematology
• Subjects: Antigens, CD34; Biological and medical sciences; Calcium - metabolism; Calcium-Calmodulin-Dependent Protein Kinase Type 1; Calcium-Calmodulin-Dependent Protein Kinases - analysis; Calcium-Calmodulin-Dependent Protein Kinases - antagonists & inhibitors; Calcium-Calmodulin-Dependent Protein Kinases - physiology; Cell Adhesion; Cell Movement; Cells, Cultured; Enzyme Inhibitors - pharmacology; Eosinophils - chemistry; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Granulocytes - chemistry; Granulocytes - physiology; Humans; Immunobiology; Myeloid cells: ontogeny, maturation, markers, receptors; Neutrophils - chemistry; Phagocytosis; Polynuclears; Respiratory Burst; RNA, Messenger - biosynthesis; Signal Transduction; Human; Immune response; Enzyme; Antibody; Transferases; Granulocyte; Leukocyte function; Activation; Chemotaxis; Eosinophil; Blood; Signal transduction; Kinase; Neutrophil; Phagocytosis
• ISSN: 0006-4971; 1528-0020
• DOI: 10.1182/blood-2004-09-3755

Activation of granulocyte effector functions, such as induction of the respiratory burst and migration, are regulated by a variety of relatively ill-defined signaling pathways. Recently, we identified a novel Ca2+/calmodulin-dependent kinase I-like kinase, CKLiK, which exhibits restricted mRNA expression to human granulocytes. Using a novel antibody generated against the C-terminus of CKLiK, CKLiK was detected in CD34+-derived neutrophils and eosinophils, as well as in mature peripheral blood granulocytes. Activation of human granulocytes by N-formyl-methionyl-leucyl-phenylalanine (fML142) and platelet-activating factor (PAF), but not the phorbol ester PMA (phorbol 12-myristate-13-acetate), resulted in induction of CKLiK activity, in parallel with a rise of intracellular Ca2+ [Ca2+]i. To study the functionality of CKLiK in human granulocytes, a cell-permeable CKLiK peptide inhibitor (CKLiK297–321) was generated which was able to inhibit kinase activity in a dose-dependent manner. The effect of this peptide was studied on specific granulocyte effector functions such as phagocytosis, respiratory burst, migration, and adhesion. Phagocytosis of Aspergillus fumigatus particles was reduced in the presence of CKLiK297–321 and fMLP-induced reactive oxygen species (ROS) production was potently inhibited by CKLiK297–321 in a dose-dependent manner. Furthermore, fMLP-induced neutrophil migration on albumin-coated surfaces was perturbed, as well as β2-integrin-mediated adhesion. These findings suggest a critical role for CKLiK in modulating chemoattractant-induced functional responses in human granulocytes.