Human -Calpain: Simple Isolation from Erythrocytes and Characterization of Autolysis Fragments
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Dusica Gabrijelcic-Geiger
Abstract
Heterodimeric calpain, consisting of the large (80 kDa) and the small (30 kDa) subunit, was isolated and purified from human erythrocytes by a highly reproducible fourstep purification procedure. Obtained material is more than 95% pure and has a specific activity of 6 7 mU/mg. Presence of contaminating proteins could not be detected by HPLC and sequence analysis. During storage at 80 C the enzyme remains fully activatable by Ca 2+ , although the small subunit is partially processed to a 22 kDa fragment. This novel autolysis product of the small subunit starts with the sequence 60 RILG and is further processed to the known 18 kDa fragment. Active forms and typical transient and stable autolysis products of the large subunit were identified by protein sequencing. In caseinzymograms only the activatable forms 80 kDa+30 kDa, 80 kDa+22 kDa and 80 kDa+18 kDa displayed caseinolysis.
Copyright © 2001 by Walter de Gruyter GmbH & Co. KG
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- Human -Calpain: Simple Isolation from Erythrocytes and Characterization of Autolysis Fragments
- Erratum
- Acknowledgement
- Content Index
- Author Index
- Subject Index
Articles in the same Issue
- New Control of Mitochondrial Membrane Potential and ROS Formation A Hypothesis
- Brix from Xenopus laevis and Brx1p From Yeast Define a New Family of Proteins Involved in the Biogenesis of Large Ribosomal Subunits
- Mig-6 Is a Negative Regulator of the Epidermal Growth Factor Receptor Signal
- β-Carotene Inhibits Growth of Human Colon Carcinoma Cells in Vitro by Induction of Apoptosis
- Ligand-Mediated Protection against Phage Lysis as a Positive Selection Strategy for the Enrichment of Epitopes Displayed on the Surface of E. coli Cells
- Structural and Redox Properties of the Leaderless DsbE (CcmG) Protein: Both Active-Site Cysteines of the Reduced Form Are Involved in Its Function in the Escherichia coli Periplasm
- Polyphenols of Cocoa: Inhibition of Mammalian 15-Lipoxygenase
- Total Antioxidant Capacity and Nuclear DNA Damage in Keratinocytes after Exposure to H2 O2
- Recombinant Cryptic Human Fibronectinase Cleaves Actin and Myosin: Substrate Specificity and Possible Role in Muscular Dystrophy
- Rat Tripeptidyl Peptidase I: Molecular Cloning, Functional Expression, Tissue Localization and Enzymatic Characterization
- Determination of NADH in Frozen Rat Brain Sections by Laser-Induced Fluorescence
- Human -Calpain: Simple Isolation from Erythrocytes and Characterization of Autolysis Fragments
- Erratum
- Acknowledgement
- Content Index
- Author Index
- Subject Index
