Related Experiment Videos
Isolation, characterization, and cloning of porcine complement component C7.
A Agah1, M C Montalto, C L Kiesecker
1Center for Experimental Therapeutics and Reperfusion Injury, Department of Anesthesiology, Perioperative and Pain Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.
Journal of Immunology (Baltimore, Md. : 1950)
|July 6, 2000
Summary
This study characterizes porcine complement component C7, revealing high structural and functional conservation with human C7. These findings advance understanding of the complement system across species.
Area of Science:
- Immunology
- Complement System Biology
Background:
- The terminal complement complex is crucial for immune responses.
- Complement component 7 (C7) is essential for terminal complement complex assembly.
- Porcine C7 had not been previously characterized.
Purpose of the Study:
- To isolate and characterize porcine complement component C7.
- To determine the structural and functional similarities between porcine and human C7.
- To investigate the expression and cross-species reactivity of porcine C7.
Main Methods:
- Affinity chromatography for porcine C7 isolation.
- Molecular cloning and sequencing of porcine C7 cDNA.
- Western blotting and mRNA expression analysis.
- Functional assays using C7-depleted human sera and cross-species reactivity tests.
Main Results:
- Porcine C7 isolated as a ~90-100 kDa glycoprotein.
- Porcine C7 shares 80% amino acid identity with human C7, including conserved structural features.
- Porcine C7 mRNA is expressed in various tissues, excluding certain leukocytes.
- Purified porcine C7 restored hemolytic activity in C7-depleted human sera.
- An inhibitory antibody against porcine C7 cross-reacted with human, rabbit, and rat C7, indicating conserved epitopes.
Conclusions:
- Porcine C7 is highly conserved structurally and functionally with human C7.
- These findings highlight conserved C7 epitopes across species, relevant for complement research.
- Characterization of porcine C7 provides a valuable model for studying complement system mechanisms.