title: A slow form of alpha-2-macroglobulin in diseased and healthy dogs.
abstract: Alpha-macroglobulins (AMs) function as non-specific protease inhibitors by using a so-called trapping mechanism, which is a compaction of the molecule that can be seen as a "fast" form in native polyacrylamide gel electrophoresis (PAGE). AMs also play a role in the transport and clearance of cytokines and growth factors from the circulation. In the dog, two AMs are known, alpha-1-macroglobulin (A1M) and alpha-2-macroglobulin (A2M). Using agarose and polyacrylamide gel electrophoresis of canine serum or plasma, we detected a cathodal, slow form of A2M. Upon activation with elastase, slow A2M resembled normal A2M in agarose gel electrophoresis, showing decreased negative charge at semi-saturation but not at full saturation with enzyme. In PAGE, however, slow A2M, unlike normal A2M, did not exhibit a "fast" form after short-term incubation with elastase. After incubation overnight, the "fast" form was seen, indicating a retarded reaction. Incubation of slow A2M with ammonium sulphate, a known activator of AMs, resulted in decreased negative charge in agarose gel electrophoresis and no reaction or partial reaction in PAGE. Slow A2M is present in fresh blood samples or may develop from partial alterations within a few days of storage. Moreover, it is sometimes reversible. Our findings may indicate that slow A2M is a result of an instability of the molecule, leading to a conformational change, which affects electrical charge and impairs the ability to develop into the "fast" form upon activation. This may lead to a delayed clearance of protease and inflammatory mediators from the circulation. Slow A2M was predominantly found in diseased dogs, especially in the Labrador retriever.
mesh_terms: D000818:Animals; D004283:Dog Diseases / Q000097:blood*; D004285:Dogs / Q000097:blood*; D004587:Electrophoresis, Agar Gel; D015180:Electrophoresis, Gel, Two-Dimensional; D010196:Pancreatic Elastase / Q000378:metabolism; D011487:Protein Conformation; D000511:alpha-Macroglobulins / Q000032:analysis* / Q000145:classification / Q000378:metabolism
chemical_list: D000511:alpha-Macroglobulins; D010196:Pancreatic Elastase