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Methods in enzymology : peptide, protein and enzyme design /
| Clasificación: | Libro Electrónico |
|---|---|
| Otros Autores: | |
| Formato: | Electrónico eBook |
| Idioma: | Inglés |
| Publicado: |
Amsterdam, Netherlands :
Academic Press,
2016.
|
| Colección: | Methods in enzymology ;
v. 580. |
| Temas: | |
| Acceso en línea: | Texto completo |
Tabla de Contenidos:
- Front Cover; Peptide, Protein and Enzyme Design; Copyright; Contents; Contributors; Preface; References; Chapter One: Chemical Posttranslational Modification with Designed Rhodium(II) Catalysts; 1. Introduction; 2. Synthesis of Rhodium(II) Conjugates as Protein Modification Catalysts; 2.1. Discussion; 2.2. Materials and General Considerations; 2.2.1. Preparation of Metalation Buffer; 2.3. Protocol 1: Preparation of Rh2(OAc)3(tfa)1; 2.4. Protocol 2: Preparation of Rhodium(II) Conjugates in Organic Solution; 2.4.1. Preparation of a Metalated FKBP Inhibitor (Coughlin et al., 2014).
- 2.5. Protocol 3: Preparation of Rhodium(II) Conjugates in Aqueous Solution2.5.1. Preparation of the Metallopeptide S2ERh; 3. Modification of an SH3 Domain and Gel-Based Visualization Thereof; 3.1. Discussion; 3.2. Materials and General Considerations; 3.2.1. Preparation of 1X Transfer Buffer; 3.2.2. Preparation of Protein Modification Buffer; 3.2.3. Preparation of MALDI-MS Matrix; 3.3. Protocol 4: Rhodium(II)-Catalyzed Protein Modification; 3.3.1. Modification of SH3 Domain of Yes Kinase in Mammalian Cell Lysate (Vohidov, Coughlin, & Ball, 2015).
- 3.4. Protocol 5: Visualization of an Alkyne-Tagged Protein by Chemical Blotting3.4.1. Fluorescent ``Chemical Blot�� Analysis of a Modified SH3 Domain; References; Chapter Two: Cell-Binding Assays for Determining the Affinity of Protein-Protein Interactions: Technologies and Considera ... ; 1. Introduction; 2. General Binding Theory and Relevance of Kd; 3. General Pitfalls in Cell-Based Binding Assays; 3.1. Time to Equilibrium; 3.2. Ligand Depletion; 4. Measuring Binding on the Surface of Yeast; 4.1. Materials; 4.1.1. Yeast Cells; 4.1.2. Solutions and Media; 4.1.3. Proteins/Antibodies.
- 4.2. Method5. Measuring Binding on the Surface of Mammalian Cells; 5.1. Direct Binding; 5.2. Competition Binding; 6. Other Methods of Measuring Binding: Kinetic Exclusion Assay and Surface Plasmon Resonance; 6.1. Kinetic Exclusion Assay; 6.2. Surface Plasmon Resonance; 6.3. Comparison; 7. Summary; Acknowledgments; References; Chapter Three: Protein and Antibody Engineering by Phage Display; 1. Introduction; 2. Equipment; 3. Materials; 3.1. Cell Lines; 3.1.1. Protocol 1: Production of SS320 Cells (Modified from Tonikian, Zhang, Boone, & Sidhu, 2007).
- 3.1.2. Protocol 2: Production of XL1-Blue Cells3.2. M13KO7 Helper Phage; 3.2.1. Protocol 3: Production of M13KO7 Helper Phage (Modified from Tonikian et al., 2007); 3.3. Phagemid Considerations; 3.4. Reagents; 3.4.1. Media; 3.4.2. Buffers (Filter Sterilize); 3.4.3. Solutions; 3.4.4. Antibiotics (Filter Sterilize); 3.4.5. Other Reagents; 4. Phage Display and Library Design; 4.1. Humanization of a Murine SUDV Antibody (Chen et al., 2014); 4.2. Mapping Hotspot Residues of Protein-Protein Interfaces (Frei et al., 2015; Stewart et al., 2013); 4.2.1. Protocol 4: Cloning into HP153.