Abstract
MRP8 and MRP14 are members of the S-100 family of Ca2+-binding proteins and are expressed by granulocytes and monocytes. Members of this family have been described to be involved in membrane and cytoskeleton interactions; we therefore studied the subcellular distribution of MRP8/MRP14 in cultured human monocytes at the ultrastructural level. Monospecific rabbit antisera against MRP8 and MRP14 and a monoclonal antibody (moAb 27E10), which exclusively recognizes the MRP8/MRP14 heterodimer but not the monomers, were used in both immunoperoxidase/preembedding-and immunogold/cryotechniques. Comparing non-stimulated monocytes with Ca2+ ionophore A23187-treated cells, we could demonstrate that MRP8 and MRP14 associate with membrane and cytoskeletal structures in a Ca2+-dependent manner. Employing moAb 27E10, MRP8/MRP14 complexes were shown to be translocated to these cellular components. In addition, immunogold double-labelling experiments revealed a clear co-localization of MRP8/MRP14 complexes with the type III intermediate filament vimentin. Analysis of immunogold-labelled cryosections of renal allografts after acute vascular rejection demonstrated that a subpopulation of infiltrating macrophages showed a similar association of MRP8/MRP14 to the cytoskeleton in situ; this finding emphasizes the in vivo relevance of our observations. We conclude that Ca2+-dependent translocation of MRP8/MRP14 occurs to distinct subcellular components suggesting a role of these proteins for the modulation of cytoskeletal and membrane interactions.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Andersson KB, Sletten K, Berner Berntzen H, Fagerhol MK, Dale I, Brandtzaeg P, Jellum E (1988) Leukocyte L1 protein and the cystic fibrosis antigen. Nature 332:688
Baudier J, Cole RD (1988) Interactions between the microtubula-associated tau proteins and S100b regulate tau phosphorylation by the Ca2+/calmodulin-dependent protein kinase II. J Biol Chem 263:5876–5883
Bhardwaj RS, Zotz C, Zwadlo-Klarwasser G, Roth J, Goebeler M, Mahnke K, Falk M, Meinardus-Hager G, Sorg C (1992) The calcium-binding proteins MRP8 and MRP14 form a membrane-associated heterodimer in a subset of monocytes/macrophages present in acute but absent in chronic inflammatory lesions. Eur J Immunol 22:1891–1897
Bianchi R, Giambanco I, Donato R (1993) S-100-protein, but not calmodulin binds glial fibrillary acidic protein and inhibits its polymerization in a clacium dependent manner. J Biol Chem 268:12669–12674
Blackwood RA, Ernst JD (1990) Characterization of Ca2+-dependent phospholipid binding, vesicle aggregation and membrane fusion by annexins. Biochem J 266:195–200
Brüggen J, Tarcsay L, Cerletti N, Odink K, Rutishauser M, Holländer G, Sorg C (1988) The molecular nature of the cystic fibrosis antigen. Nature 331:570
Chou Y-H, Rosevear E, Goldman RD (1989) Phosphorylation and disassembly of intermediate filaments in mitotic cells. Proc Natl Acad Sci USA 86:1885–1889
Dale I, Brandtzaeg P, Fagerhol MK, Scott H (1985) Distribution of a new myelomonocytic antigen (L1) in human peripheral blood leukocytes. Immunofluorescence and immunoperoxidase staining features in comparison with lysozyme and lactoferrin. Am J Clin Pathol 84:24–34
Donato R (1991) Perspectives in S-100 protein biology. Cell Calcium 12:713–726
Dorin JR, Novak M, Hill RE, Brock DJH, Secher DS, Heyningen V van (1987) A clue to the basic defect in cystic fibrosis from cloning the CF antigen gene. Nature 326:614–617
Edgeworth J, Freemont P, Hogg N (1989) Ionomycin-regulated phosphorylation of the myeloid calcium-binding protein p 14. Nature 342:189–192
Glenney JR Jr (1987) Calpactins: calcium-regulated membrane-skeletal proteins. Bio Essays 7:173–175
Goebeler M, Roth J, Henseleit U, Sunderkötter C, Sorg C (1993) Expression and complex assembly of calcium-binding proteins MRP8 and MRP14 during differentiation of murine myelomonocytic cells. J Leukocyte Biol 53:11–18
Hessian PA, Edgeworth J, Hogg N (1993) MRP8 and MRP14, two abundant calcium-binding proteins of neutrophils and monocytes. Leukocyte Biol 53:197–204
Hogg N, Allen C, Edgeworth J (1989) Monoclonal antibody 5.5 reacts with p8, 14, a myeloid molecule associated with some vascular endothelium. Eur J Immunol 19:1053–1061
Kligman D, Hilt DC (1991) The S-100 protein family: a biochemical and functional overview. In: Heizmann CW (ed) Novel Ca2+-binding proteins. Springer, Berlin Heidelberg New York, pp 65–103
Langanger G, DeMay J (1989) Detection of cytoskeletal proteins in cultured cells at the ultrastructural level. In: Verkleii AJ, Leunissen JLM (eds) Immuno-gold labelling in cell biology. CRC Press Boca Raton, Fla., pp 335–351
Lemarchand P, Vaglio M, Mauël J, Markert M (1992) Translocation of a small cytosolic calcium-binding protein (MRP-8) to plasma membrane correlates with human neutrophil activation. J Biol Chem 267:19379–19382
Luft JH (1961) Improvements in epoxy restin embedding methods. J Biophys Biochem Cytol 9:409–414
McLean IW, Nakane PK (1974) Periodate-lysine-paraform-aldehyde fixative. A new fixative for immunoelectron microscopy. J Histochem Cytochem 22:1077–1083
Murao S, Collart FR, Huberman E (1989) A protein containing the cystic fibrosis antigen is an inhibitor of protein kinases. J Biol Chem 264:8356–8360
Odink K, Cerletti N, Brüggen J, Clerc RG, Tarcsay L, Zwadlo G, Gerhards G, Schlegel R, Sorg C (1987) Two calcium-binding proteins in infiltrate macrophages of rheumatoid arthritis. Nature 330:80–82
Roth J, Goebeler M, Bos C van den, Sorg C (1993a) Expression of calcium-binding proteins MRP8 and MRP14 is associated with distinct monocytic differentiation pathways in HL-60 cells. Biochem Biophys Res Commun 191:565–570
Roth J, Burwinkel F, Bos C van den, Goebeler M, Vollmer E, Sorg C (1993b) MRP8 and MRP14, S-100-like proteins associated with myeloid differentiation, are translocated to plasma membrane and intermediate filaments in a calcium-dependent manner. Blood 82:1875–1883
Selinfreund RH, Barger SW, Welsh MJ, Van Eldik LJ (1990) Antisense inhibition of glial S100β production results in alterations in cell morphology, cytoskeletal organization, and cell proliferation. J Cell Biol 111:2021–2028
Sitte H, Neumann K, Edelmann L (1986) Cryofixation and cryosubstitution for routine work in transmission electron microscopy. In: Müller M, Becker RP, Bovde A, Wolosewick JJ (SEM) The science of biological specimen preparation. SEM O'Hare, Chicago, pp 103–118
Teigelkamp S, Bhardwaj RS, Roth J, Meinardus-Hager G, Karas M, Sorg C (1991) Calcium-dependent complex assembly of the myeloic differentiation proteins MRP-8 and MRP-14. J Biol Chem 266:13462–13467
Wilkinson MM, Busuttil A, Hayward C, Brock DJH, Dorin JR, Heyningen V van (1988) Expression pattern of two related cystic fibrosis-associated calcium-binding proteins in normal and abnormal tissues. J Cell Sci 91:221–230
Zwadlo G, Schlegel R, Sorg C (1986) A monoclonal antibody to a subset of human monocytes found only in the peripheral blood and inflammatory tissues. J Immunol 137:512–518
Zwadlo G, Brüggen J, Gerhards G, Schlegel R, Sorg C (1988) Two calcium-binding proteins associated with specific stages of myeloid cell differentiation are expressed by subsets of macrophages in inflammatory tissues. Clin Exp Immunol 72:510–515
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Burwinkel, F., Bitter, U., Vollmer, E. et al. Ultrastructural localization of the S-100-like proteins MRP8 and MRP14 in monocytes is calcium-dependent. Histochemistry 101, 113–120 (1994). https://doi.org/10.1007/BF00269357
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00269357