Calpastatin Exon 1B-Derived Peptide, a Selective Inhibitor of Calpain: Enhancing Cell Permeability by Conjugation with Penetratin
-
S. Gil-Parrado
Abstract
The ubiquitous calpains, u- and m-calpain, have been implicated in essential physiological processes and various pathologies. Cell-permeable specific inhibitors are important tools to elucidate the roles of calpains in cultivated cells and animal models. The synthetic Nacetylated 27-mer peptide derived from exon B of the inhibitory domain 1 of human calpastatin (CP1B) is unique as a potent and highly selective reversible calpain inhibitor, but is poorly cell-permeant. By addition of N-terminal cysteine residues we have generated a disulfide-conjugated CP1B with the cell-penetrating 16-mer peptide penetratin derived from the third helix of the Antennapedia homeodomain protein. The inhibitory potency and selectivity of CP1B for calpain versus cathepsin B and L, caspase 3 and the proteasome was not affected by the conjugation with penetratin. The conjugate was shown to efficiently penetrate into living LCLC 103H cells, since it prevents ionomycin-induced calpain activation at 200-fold lower concentration than the non-conjugated inhibitor and is able to reduce calpain triggered apoptosis of these cells. Penetratin-conjugated CP1B seems to be a promising alternative to the widely used cell-permeable peptide aldehydes (e.g. calpain inhibitor I) which inhibit the lysosomal cathepsins and partially the proteasome as well or even better than the calpains.
Copyright © 2003 by Walter de Gruyter GmbH & Co. KG
Articles in the same Issue
- Roumen Tsanev, Pioneer of the Early Days of Nucleic Acid Gel Electrophoresis and Histone Code Epigenetics
- Recombinant RNase Z Does Not Recognize CCA as Part of the tRNA and Its Cleavage Efficieny Is Influenced by Acceptor Stem Length
- HIV-1 TAR as Anchoring Site for Optimized Catalytic RNAs
- Transcription of Different Exons 1 of the Human Neuronal Nitric Oxide Synthase Gene Is Dynamically Regulated in a Cell- and Stimulus- Specific Manner
- Hepatitis B Virus Core Antigen: Enhancement of Its Production in Escherichia coli, and Interaction of the Core Particles with the Viral Surface Antigen
- Determinants of Enzymatic Specificity in the Cys-Met-Metabolism PLP-Dependent Enzyme Family: Crystal Structure of Cystathionine γ-Lyase from Yeast and Intrafamiliar Structure Comparison
- In vitro Uptake and Stability Study of pVEC and Its All-D Analog
- Calpastatin Exon 1B-Derived Peptide, a Selective Inhibitor of Calpain: Enhancing Cell Permeability by Conjugation with Penetratin
- Structure-Activity Relationships of Glucose-Dependent Insulinotropic Polypeptide (GIP)
- Interleukin-6 N-Terminal Peptides Modulate the Expression of junB Protooncogene and the Production of Fibrinogen in HepG2 Cells
- Induction of Apoptosis in Vascular Cells by Plasminogen Activator Inhibitor-1 and High Molecular Weight Kininogen Correlates with Their Anti-Adhesive Properties
- The Role of Octadecanoids and Functional Mimics in Soybean Defense Responses
- Invasiveness of Transformed Human Breast Epithelial Cell Lines Is Related to Cathepsin B and Inhibited by Cysteine Proteinase Inhibitors
- Retinoids Inhibit Human Epidermal Keratinocyte RNase P Activity
- A Putative Glutathione Peroxidase of Drosophila Encodes a Thioredoxin Peroxidase That Provides Resistance against Oxidative Stress But Fails to Complement a Lack of Catalase Activity
- Characterization of Calmodulin Binding to the Orphan Nuclear Receptor ERRγ
- Evidence for Coordinated Induction and Repression of Ecto-5'-Nucleotidase (CD73) and the A2a Adenosine Receptor in a Human B Cell Line
- Bauhinia Proteinase Inhibitor-Based Synthetic Fluorogenic Substrates for Enzymes Isolated from Insect Midgut and Caterpillar Bristles
- Synthesis and Characterization of EMPO-Derived 5,5-Disubstituted 1-Pyrroline N-Oxides as Spin Traps Forming Exceptionally Stable Superoxide Spin Adducts
Articles in the same Issue
- Roumen Tsanev, Pioneer of the Early Days of Nucleic Acid Gel Electrophoresis and Histone Code Epigenetics
- Recombinant RNase Z Does Not Recognize CCA as Part of the tRNA and Its Cleavage Efficieny Is Influenced by Acceptor Stem Length
- HIV-1 TAR as Anchoring Site for Optimized Catalytic RNAs
- Transcription of Different Exons 1 of the Human Neuronal Nitric Oxide Synthase Gene Is Dynamically Regulated in a Cell- and Stimulus- Specific Manner
- Hepatitis B Virus Core Antigen: Enhancement of Its Production in Escherichia coli, and Interaction of the Core Particles with the Viral Surface Antigen
- Determinants of Enzymatic Specificity in the Cys-Met-Metabolism PLP-Dependent Enzyme Family: Crystal Structure of Cystathionine γ-Lyase from Yeast and Intrafamiliar Structure Comparison
- In vitro Uptake and Stability Study of pVEC and Its All-D Analog
- Calpastatin Exon 1B-Derived Peptide, a Selective Inhibitor of Calpain: Enhancing Cell Permeability by Conjugation with Penetratin
- Structure-Activity Relationships of Glucose-Dependent Insulinotropic Polypeptide (GIP)
- Interleukin-6 N-Terminal Peptides Modulate the Expression of junB Protooncogene and the Production of Fibrinogen in HepG2 Cells
- Induction of Apoptosis in Vascular Cells by Plasminogen Activator Inhibitor-1 and High Molecular Weight Kininogen Correlates with Their Anti-Adhesive Properties
- The Role of Octadecanoids and Functional Mimics in Soybean Defense Responses
- Invasiveness of Transformed Human Breast Epithelial Cell Lines Is Related to Cathepsin B and Inhibited by Cysteine Proteinase Inhibitors
- Retinoids Inhibit Human Epidermal Keratinocyte RNase P Activity
- A Putative Glutathione Peroxidase of Drosophila Encodes a Thioredoxin Peroxidase That Provides Resistance against Oxidative Stress But Fails to Complement a Lack of Catalase Activity
- Characterization of Calmodulin Binding to the Orphan Nuclear Receptor ERRγ
- Evidence for Coordinated Induction and Repression of Ecto-5'-Nucleotidase (CD73) and the A2a Adenosine Receptor in a Human B Cell Line
- Bauhinia Proteinase Inhibitor-Based Synthetic Fluorogenic Substrates for Enzymes Isolated from Insect Midgut and Caterpillar Bristles
- Synthesis and Characterization of EMPO-Derived 5,5-Disubstituted 1-Pyrroline N-Oxides as Spin Traps Forming Exceptionally Stable Superoxide Spin Adducts
