Methods in enzymology : peptide, protein and enzyme design / edited by Vincent L. Pecoraro, Department of Chemistry University of Michigan, Ann Arbor MI, United States.

Format:

Book

Contributor:

Pecoraro, Vincent L., editor.

Series:

Methods in Enzymology

Methods in Enzymology ; v.Volume 580

Language:

English

Subjects (All):

Enzymes.

Physical Description:

1 online resource (686 p.)

Edition:

1st ed.

Place of Publication:

Amsterdam, Netherlands : Academic Press, 2016.

Summary:

De Novo Enzyme Design, the newest volume in the Methods in Enzymology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field.

Contents:

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

4.2.2. Protocol 5: Amplifying and Titering Phage

Notes:

Includes indexes.

Description based on online resource; title from PDF title page (ebrary, viewed September 12, 2016).

Description based on publisher supplied metadata and other sources.

ISBN:

9780128054345

0128054344

9780128053805

0128053801

OCLC:

957614433