Purification and partial characterization of α1-proteinase inhibitor in the common marmoset (Callithrix jacchus).
Fecal alpha1-proteinase inhibitor (α1-PI) concentration has been to diagnose enteric protein loss in dogs and cats. Chronic lymphocytic enteritis is commonly seen in the marmoset (Callithrix jaccus) and is characterized by hypoalbuminemia. As a prelude to immunoassay development for detecting enteric protein loss, marmoset serum α1-PI was purified using immunoaffinity chromatography and ceramic hydroxyapatite chromatography. Partial characterization was performed by reducing gel electrophoresis and enzyme inhibitory assays. Protein identity was confirmed with peptide mass fingerprinting and N-terminal amino acid sequencing. Molecular mass, relative molecular mass, and isoelectric point for marmoset α1-PI were 54 kDa, 51,677, and 4.8-5.4, respectively. Trypsin, chymotrypsin, and elastase inhibitory activity were observed. N-terminal amino acid sequence for marmoset α1-PI was EDPQGDAAQKMDTSHH. In conclusion, marmoset α1-PI was successfully purified from serum with an overall yield of 12% using a rapid and efficient method. Purified marmoset α1-PI has characteristics similar to those of α1-PI reported for other species.
Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, College of Veterinary and Biomedical Sciences, Texas A&M University, College Station, Texas 77843-4474. Electronic address: JCyrus@cvm.tamu.edu.
Joseph Cyrus
JC
Parambeth
Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, College of Veterinary and Biomedical Sciences, Texas A&M University, College Station, Texas 77843-4474.
Jan S
JS
Suchodolski
Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, College of Veterinary and Biomedical Sciences, Texas A&M University, College Station, Texas 77843-4474.
Jörg M
JM
Steiner
D000818:Animals; D002144:Callithrix / Q000235:genetics* / Q000378:metabolism; D002846:Chromatography, Affinity / Q000662:veterinary; D002918:Chymotrypsin / Q000378:metabolism; D004751:Enteritis / Q000150:complications / Q000662:veterinary*; D034141:Hypoalbuminemia / Q000209:etiology / Q000662:veterinary*; D007526:Isoelectric Point; D008970:Molecular Weight; D010196:Pancreatic Elastase / Q000378:metabolism; D010449:Peptide Mapping; D020539:Sequence Analysis, Protein; D014357:Trypsin / Q000378:metabolism; D000515:alpha 1-Antitrypsin / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
D000515:alpha 1-Antitrypsin; D002918:Chymotrypsin; D010196:Pancreatic Elastase; D014357:Trypsin
Characterization; Common marmoset; Purification; alpha(1)-proteinase inhibitor