Ionomycin-regulated phosphorylation of the myeloid calcium-binding protein p14.
Two associated calcium-binding proteins (CaBPs) have recently been identified specifically in cells of myeloid origin. These proteins have relative molecular masses (Mr) of 8,000 and 14,000 and are variously referred to as the cystic fibrosis antigen, the L1 light chain, MRP-8 or p8, and the L1 heavy chain, MRP14 or p14, respectively. The expression of p8 and p14 seems to be confined to a specific stage of myeloid cell differentiation, because both proteins are expressed in circulating neutrophils and monocytes but not in normal tissue macrophages. In chronic inflammatory conditions, however, such as rheumatoid arthritis, macrophages in affected tissues express both p8 and p14. These proteins are members of a family of CaBPs of low Mr, which include S-100 alpha and beta proteins, calcyclin (2A9), intestinal CaBP and p11. All the proteins have an Mr of approximately 10,000 with the exception of p14 which has a longer C-terminal sequence after the second calcium-binding domain. Little is known about their function, although by analogy with calmodulin they could be molecules involved in intracellular signalling that are activated by an increase in the intracellular Ca2+ concentration ([Ca2+]). Here we report that p14 is phosphorylated in both monocytes and neutrophils. The level of p14 phosphorylation can be increased by elevating the [Ca2+]i using the ionophore ionomycin, but is not affected by activation of protein kinase C using phorbol 12,13-dibutyrate. The phosphorylated residue is threonine at position 113, which is the penultimate amino acid in p14 and contained in the longer 'tail' sequence. Part of this sequence is identical to the neutrophil immobilizing factors NIF-1 and NIF-2, indicating that the phosphorylation event could have a role in the generation of NIF activity in the p14 protein.
Edgeworth
J
J
Macrophage Laboratory, Lincoln's Inn Fields, UK.
Freemont
P
P
Hogg
N
N
D000595:Amino Acid Sequence; D002118:Calcium / Q000502:physiology; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D015759:Ionomycin / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009000:Monocytes / Q000502:physiology*; D009504:Neutrophils / Q000502:physiology*; D015240:Phorbol 12,13-Dibutyrate / Q000494:pharmacology; D010766:Phosphorylation; D019000:Phosphotyrosine; D015398:Signal Transduction; D014443:Tyrosine / Q000378:metabolism
D002135:Calcium-Binding Proteins; D019000:Phosphotyrosine; D015240:Phorbol 12,13-Dibutyrate; D014443:Tyrosine; D015759:Ionomycin; D002118:Calcium