ABSTRACT
Three related alpha-protease inhibitors, PI2 I, PI3 C and PI4 C2, of blood serum of the pig (Sus scrofa) were isolated. PI2 I inhibited both trypsin and chymotrypsin; PI3 C and PI4 C2 strongly inhibited chymotrypsin, but did not significantly inhibit trypsin. By using SDS-PAGE, the three proteins were found to be composed of single polypeptide chains, and molecular weights were 63,000 for PI2 I, 58,000 for PI3 C and 64,000 for PI4 C2. All three proteins were shown to be glycoproteins. In PI3 C, eight sialic acid residues were found, and in PI4 C2 (similarly as in PI2 F) 10-11 residues were found. Amino acid composition as well as N-terminal sequences of the three proteins were very similar, indicating close homology. Comparison of these partial amino acid sequences with the cDNA-deduced amino acid sequence of pig alpha-antichymotrypsin (AACT; Buchman, 1989, GenBank, Accession No. M29508) revealed great similarities, the sequence of PI2 I being virtually identical with the pig AACT. On the basis of all available results, PI2 is proposed to be pig AACT, an orthologue of human AACT.
Subject(s)
Blood Proteins/chemistry , Chymotrypsin/antagonists & inhibitors , Protease Inhibitors/blood , alpha 1-Antichymotrypsin/blood , Amino Acid Sequence , Amino Acids/analysis , Animals , DNA, Complementary/chemistry , Hydrogen-Ion Concentration , Molecular Sequence Data , Molecular Weight , Neuraminidase/pharmacology , Sequence Homology , Swine , Trypsin Inhibitors/blood , alpha 1-Antichymotrypsin/chemistryABSTRACT
A further alpha-protease inhibitor system, PI4, was detected in porcine sera using either 2D agarose gel, pH 5.0-PAGE, pH 9.0, or 1D PAGE followed by immunoblotting with rabbit anti-porcine PI2 or PI3 antisera. PI4 inhibited chymotrypsin, but not trypsin. Seven allelic variants of PI4 were described. By haplotyping of alpha-protease inhibitor systems in 52 complete families it was shown that PI4 locus belongs to the PI gene cluster. The probable order of the PI loci was: PI1, PO1A, PI2, PI4, PI3.
Subject(s)
Blood Proteins/genetics , Multigene Family , Protease Inhibitors , Swine/genetics , Animals , Electrophoresis, Agar Gel/veterinary , Electrophoresis, Gel, Two-Dimensional/veterinary , Haplotypes , Immunoblotting/veterinaryABSTRACT
1. Serum proteins of Equus grevyi, E. zebra hartmannae, E. burchelli boehmi, E. b. chapmanni and E. b. antiquorum were studied using starch-gel electrophoresis, 1-D polyacrylamide-gel electrophoresis, inhibitions of trypsin and chymotrypsin, immunoblotting, and specific staining for esterase. 2. Clear species-specific patterns were observed in albumin, transferrin, and for E. grevyi in protease inhibitor-1. Specific esterase was detected only in E. z. hartmannae. 3. Protein polymorphism was found in all studied species: E. grevyi--transferrin; E. z. hartmannae--protease inhibitor-1; E. b. boehmi--albumin, GC, transferrin, protease inhibitor-1, protease inhibitor-T; E. b. chapmanni--albumin, GC, transferrin, protease inhibitor-1; E. b. antiquorum--GC, transferrin, protease inhibitor-1. 4. Phenotype patterns of the polymorphic proteins were indicative of simple codominant inheritance. Further studies of polymorphism of protease inhibitor-2 and variability of protease inhibitor-X are needed. 5. alpha 1B glycoprotein in all zebra species was monomorphic. 6. The main transferrin components and alpha 1B glycoprotein of zebra (E. b. boehmi) were characterized for terminal sialic acid content.
Subject(s)
Blood Proteins/isolation & purification , Perissodactyla/blood , Animals , Blood Protein Electrophoresis , Blood Proteins/genetics , Esterases/blood , Esterases/genetics , Genetic Variation , Perissodactyla/classification , Perissodactyla/genetics , Polymorphism, Genetic , Protease Inhibitors/blood , Serum Albumin/genetics , Serum Albumin/isolation & purification , Species Specificity , Transferrin/genetics , Transferrin/isolation & purification , Vitamin D-Binding Protein/genetics , Vitamin D-Binding Protein/isolation & purificationABSTRACT
Linkage relationships between A1BG, PGD, and HPX loci of rabbits were studied on segregation data in backcross matings. No sign of linkage between the three studied loci was found.
Subject(s)
Blood Proteins/genetics , Genetic Linkage/genetics , Hemopexin/genetics , Phosphogluconate Dehydrogenase/genetics , Rabbits/genetics , Alleles , Animals , Crosses, GeneticABSTRACT
Various electrophoretic techniques, immunoblotting and inhibitions of trypsin and chymotrypsin were used to study the variability of serum proteins in farmed red deer, Cervus elaphus L., of Czechoslovakian origin. Easily interpretable polymorphisms were observed in transferrin (variants A, B1, B2, C) and vitamin D binding protein, GC (variants D, F, I, S). Great variability was observed in the protease inhibitors PI2, PI3, PI4, PI5, and PI8 and in unidentified zones in the vicinity of albumin, but no genetical or physiological interpretation for this variability is yet available. Haemopexin, alpha 1 glycoprotein, protease inhibitors PI1, PI6 and PI7 were monomorphic.
