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Expression of 67,000 MW calcimedin and its binding protein in resident and thioglycolate-elicited macrophages
1Rockefeller University, Laboratory Animal Research Center, New York, New York 10021.
Abstract:
Calcimedins are newly recognized calcium-dependent hydrophobic-binding proteins. The 67,000 MW calcimedin exhibits calcium-dependent specific binding to a 60,000 MW intracellular acceptor protein (60,000 MW). Macrophages possessed 30,000 MW and 67,000 MW calcimedins and 60,000 MW acceptor protein, identified by immunofluorescence and confirmed by SDS-PAGE and Western immunoblots. Compared with low levels in resident macrophages, thioglycolate-elicited inflammatory macrophages showed large increases in the levels of 67,000 MW calcimedin and 60,000 MW acceptor protein, especially at cell-limiting membranes. Levels of calmodulin and 30,000 MW calcimedin appeared similar in both populations. Trypsinization removed most 67,000 MW calcimedin and some 60,000 MW acceptor-protein staining. The calcimedins appear to represent novel macrophage-differentiation antigens. Their modulation suggests that they may play a role in the inflammatory response.
Insights
Newly discovered calcimedin proteins bind calcium and hydrophobic molecules. Their increased levels in inflammatory macrophages suggest a role in the immune response.
Area of Science:
- Immunology
- Cell Biology
- Protein Biochemistry
Background:
- Calcimedins are newly identified calcium-dependent hydrophobic-binding proteins.
- These proteins interact with intracellular acceptor proteins, influencing cellular functions.
Purpose of the Study:
- To investigate the presence and modulation of calcimedins and their acceptor proteins in macrophages.
- To determine the potential role of calcimedins in the inflammatory response.
Main Methods:
- Immunofluorescence staining to identify protein localization in macrophages.
- SDS-PAGE and Western immunoblots for protein confirmation and quantification.
- Analysis of protein levels in resident versus elicited inflammatory macrophages.
Main Results:
- Macrophages express 30,000 MW and 67,000 MW calcimedins and a 60,000 MW acceptor protein.
- Elicited inflammatory macrophages show significantly increased levels of 67,000 MW calcimedin and 60,000 MW acceptor protein compared to resident macrophages.
- Trypsinization partially removed these proteins, indicating surface expression or accessibility.
Conclusions:
- Calcimedins and their acceptor protein are modulated during macrophage inflammation.
- These proteins may function as novel macrophage-differentiation antigens.
- The observed modulation suggests a role for calcimedins in the macrophage inflammatory response.