Fractionation and partial characterization of alpha-1-protease isoinhibitors of horse.
Abstract: The principal alpha-1-protease inhibitor of horse was fractionated by classical methods and analysed with a modified fibrinogen-agarose gel electrophoretic method of high sensitivity and resolving power. Starting with an electrophoretically homogeneous inhibitor in unfractionated serum, two isoinhibitor bands became apparent after fractionation with (NH4)2SO4 and DEAE-cellulose DE-52 ion-exchange chromatography. The isoinhibitors differed in electrophoretic migration and in the elution pattern from Sephadex G-100 gel filtration, but possessed identical antigenic determinants and enzyme specificity. The slower migrating isoinhibitor with an apparent molecular weight of 90 000 could be highly purified. In contrast the faster moving isoinhibitor (molecular weight 65 000) could not be completely freed from a contaminating alpha-2-protease inhibitor. The formation of the two isoinhibitors is discussed considering conformational changes analogous to phenomena observed with alpha-2-macroglobulin, or dimer formation in combination with altered conformations. The isoinhibitors described here are new additions to the different heterogeneities which exist in alpha-1-protease inhibitors in horse. They also supplement the different heterogeneities which exist among the alpha-1-protease inhibitors of mammals.
Publication Date: 1980-12-04 PubMed ID: 6971123DOI: 10.1016/0005-2744(80)90152-7Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This study focuses on classifying and analyzing a vital alpha-1-protease inhibitor in horses. The researchers find variants of the inhibitor that differ in molecular weights but have similar properties.
Research Overview
The primary intention of the research was to identify, separate, and characterize the alpha-1-protease inhibitors found within horses.
- The research began from an alpha-1-protease inhibitor in a homogeneous horse serum.
- There was the application of standard separation techniques leading to the emergence of two separate isoinhibitor bands.
- The distinction of these bands was based on differing molecular weights and elution patterns.
Separation Techniques
Classical methods involving ammonium sulphate ((NH4)2SO4) and DEAE-cellulose DE-52 ion-exchange chromatography were used to separate the isoinhibitors.
- The (NH4)2SO4 technique is a well-known preliminary fractionation method in protein purification.
- The DEAE-cellulose DE-52 ion-exchange chromatography aids in the isolation of these proteins based on the net charge.
Characterization of Isoinhibitors
Properties of Isoinhibitors
- Both bands displayed identical antigenic determinants i.e., same areas on the antigen molecule that antibodies or cells can recognize and respond to.
- They demonstrated equivalent enzyme specificities, meaning they bind to their respective enzymes in the same manner.
Difference in Molecular Weight
- The isoinhibitor with a higher molecular weight (90,000) showed slower migration but could also be highly purified.
- The one with a lower molecular weight (65,000), though it moved faster, could not be completely separated from an alpha-2-protease inhibitor.
Derivation of Isoinhibitors
The researchers hypothesize that the process of formation of these isoinhibitors could be correlated to the phenomena observed with alpha-2-macroglobulin, or due to dimer formation along with altered conformations.
Expansion of Knowledge
The discovery of these isoinhibitors contributes to the growing understanding of heterogeneities existing within the alpha-1-protease inhibitors in horses, as well as their counterparts in other mammals.
Cite This Article
APA
Pellegrini A, von Fellenberg R.
(1980).
Fractionation and partial characterization of alpha-1-protease isoinhibitors of horse.
Biochim Biophys Acta, 616(2), 351-361.
https://doi.org/10.1016/0005-2744(80)90152-7 Publication
Researcher Affiliations
MeSH Terms
- Animals
- Blood Proteins
- Chromatography
- Horses
- Immunochemistry
- Isoelectric Focusing
- Molecular Conformation
- Molecular Weight
- Protease Inhibitors / blood
- Protease Inhibitors / immunology
- Proteins / immunology
- alpha 1-Antitrypsin
Citations
This article has been cited 3 times.- Kuehn L, Rutschmann M, Dahlmann B, Reinauer H. Proteinase inhibitors in rat serum. Purification and partial characterization of three functionally distinct trypsin inhibitors. Biochem J 1984 Mar 15;218(3):953-9.
- Potempa J, Wunderlich JK, Travis J. Comparative properties of three functionally different but structurally related serpin variants from horse plasma. Biochem J 1991 Mar 1;274 ( Pt 2)(Pt 2):465-71.
- Patterson SD, Bell K, Shaw DC. The equine major plasma serpin multigene family: partial characterization including sequence of the reactive-site regions. Biochem Genet 1991 Oct;29(9-10):477-99.
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