Note: Zugl.: Karlsruhe, Karlsruher Inst. für Technologie, Diss., 2011 , Systemvoraussetzungen: Acrobat reader.

Note: Cover -- Title Page -- Copyright -- Contents -- Preface -- Chapter 1 Characterization of Protein Molecules Prepared by Total Chemical Synthesis -- 1.1 Introduction -- 1.2 Chemical Protein Synthesis -- 1.3 Comments on Characterization of Synthetic Protein Molecules -- 1.3.1 Homogeneity -- 1.3.2 Amino Acid Sequence -- 1.3.3 Chemical Analogues -- 1.3.4 Limitations of SPPS -- 1.3.5 Folding as a Purification Step -- 1.4 Summary -- References -- Chapter 2 Automated Fast Flow Peptide Synthesis -- 2.1 Introduction -- 2.2 Results -- 2.2.1 Summary -- 2.2.1.1 Mechanical Principles -- 2.2.1.2 Chemical Principles -- 2.2.1.3 User Interface Principles -- 2.2.1.4 Data Analysis Method -- 2.2.1.5 Outcome -- 2.2.2 First‐generation Automated Fast Flow Peptide Synthesis -- 2.2.2.1 Key Findings -- 2.2.2.2 Design of First‐generation AFPS -- 2.2.2.3 Characterization of First‐generation AFPS -- 2.2.3 Second‐generation Automated Fast Flow Peptide Synthesis -- 2.2.3.1 Key Findings -- 2.2.3.2 Design of Second‐generation AFPS -- 2.2.3.3 Characterization and Use of Second‐generation AFPS -- 2.2.4 Third‐generation Automated Fast Flow Peptide Synthesis -- 2.2.4.1 Key Findings -- 2.2.4.2 Design of Third‐generation AFPS -- 2.2.4.3 Characterization of Third‐generation AFPS -- 2.2.4.4 Reagent Stability Study -- 2.2.5 Fourth‐generation Automated Fast Flow Peptide Synthesis -- 2.2.5.1 Key Findings -- 2.2.5.2 Effect of Solvent on Fast Flow Synthesis -- 2.2.5.3 Design and Characterization of Fourth‐generation AFPS -- 2.3 Conclusions -- Acknowledgments -- References -- Chapter 3 N,S‐ and N,Se‐Acyl Transfer Devices in Protein Synthesis -- 3.1 Introduction -- 3.2 N,S‐ and N,Se‐Acyl Transfer Devices: General Presentation, Reactivity and Statistical Overview of Their Utilization -- 3.2.1 General Presentation of N,S‐ and N,Se‐Acyl Transfer Devices. , 3.2.2 Relative Reactivity of N,S‐ and N,Se‐Acyl Transfer Devices -- 3.2.3 A Statistical Overview of the Synthetic Use of N,S‐ and N,Se‐Acyl Transfer Devices for Protein Total Chemical Synthesis -- 3.3 Preparation of SEA/SeEAoff and SEAlide Peptides -- 3.3.1 Preparation of SEA and SeEA Peptides -- 3.3.2 Preparation of SEAlide Peptides -- 3.4 Redox‐controlled Assembly of Biotinylated NK1 Domain of the Hepatocyte Growth Factor (HGF) Using SEA and SeEA Chemistries -- 3.5 The Total Chemical Synthesis of GM2‐AP Using SEAlide‐based Chemistry -- 3.6 Conclusion -- References -- Chapter 4 Chemical Synthesis of Proteins Through Native Chemical Ligation of Peptide Hydrazides -- 4.1 Introduction -- 4.2 Development of Peptide Hydrazide‐based Native Chemical Ligation -- 4.2.1 Conversion of Peptide Hydrazide to Peptide Azide -- 4.2.2 Acyl Azide‐based Solid‐phase Peptide Synthesis -- 4.2.3 Acyl Azide‐based Solution‐phase Peptide Synthesis -- 4.2.4 The Transesterification of Acyl Azide -- 4.2.5 Development of Peptide Hydrazide‐based Native Chemical Ligation -- 4.3 Optimization of Peptide Hydrazide‐based Native Chemical Ligation -- 4.3.1 Preparation of Peptide Hydrazides -- 4.3.1.1 2‐Cl‐Trt‐Cl Resin -- 4.3.1.2 Peptide Hydrazides from Expressed Proteins -- 4.3.1.3 Sortase‐mediated Hydrazide Generation -- 4.3.2 Activation Methods of Peptide Hydrazide -- 4.3.2.1 Knorr Pyrazole Synthesis -- 4.3.2.2 Activation in TFA -- 4.3.3 Ligation Sites of Peptide Hydrazide -- 4.3.4 Multiple Fragment Ligation Based on Peptide Hydrazide -- 4.3.4.1 N‐to‐C Sequential Ligation -- 4.3.4.2 Convergent Ligation -- 4.3.4.3 One‐pot Ligation -- 4.4 Application of Peptide Hydrazide‐based Native Chemical Ligation -- 4.4.1 Peptide Drugs and Diagnostic Tools -- 4.4.1.1 Peptide Hydrazides for Cyclic Peptide Synthesis -- 4.4.1.2 Screening for d Peptide Inhibitors Targeting PD‐L1. , 4.4.1.3 Chemical Synthesis of DCAF for Targeted Antibody Blocking -- 4.4.1.4 Peptide Toxins -- 4.4.2 Synthesis and Application of Two‐photon Activatable Chemokine CCL5 -- 4.4.3 Proteins with Posttranslational Modification -- 4.4.3.1 The Synthesis of Glycosylation‐modified Full‐length IL‐6 -- 4.4.3.2 The Chemical Synthesis of EPO -- 4.4.3.3 Chemical Synthesis of Homogeneous Phosphorylated p62 -- 4.4.3.4 Chemical Synthesis of K19, K48 Bi‐acetylated Atg3 Protein -- 4.4.4 Ubiquitin Chains -- 4.4.4.1 Synthesis of K27‐linked Ubiquitin Chains -- 4.4.4.2 Synthesis of Atypical Ubiquitin Chains by Using an Isopeptide‐linked Ub Isomer -- 4.4.4.3 Synthesis of Atypical Ubiquitin Chains Using an Isopeptide‐linked Ub Isomer -- 4.4.5 Modified Nucleosomes -- 4.4.5.1 Synthesis of DNA‐barcoded Modified Nucleosome Library -- 4.4.5.2 Synthesis of Modified Histone Analogs with a Cysteine Aminoethylation‐assisted Chemical Ubiquitination Strategy -- 4.4.5.3 Synthesis of Ubiquitylated Histones for Examination of the Deubiquitination Specificity of USP51 -- 4.4.6 Membrane Proteins -- 4.4.7 Mirror‐image Biological Systems -- 4.5 Summary and Outlook -- References -- Chapter 5 Expanding Native Chemical Ligation Methodology with Synthetic Amino Acid Derivatives -- 5.1 Native Chemical Ligation -- 5.2 Desulfurization Chemistries -- 5.3 Aspartic Acid (Asp, D) -- 5.4 Glutamic Acid (Glu, E) -- 5.5 Phenylalanine (Phe, F) -- 5.6 Isoleucine (Ile, I) -- 5.7 Lysine (Lys, K) -- 5.8 Leucine (Leu, L) -- 5.9 Asparagine (Asn, N) -- 5.10 Proline (Pro, P) -- 5.11 Glutamine (Gln, Q) -- 5.12 Arginine (Arg, R) -- 5.13 Threonine (Thr, T) -- 5.14 Valine (Val, V) -- 5.15 Tryptophan (Trp, W) -- 5.16 Application of Selenocysteine (Sec) to Ligation Chemistry -- 5.17 Aspartic Acid (Asp, D) -- 5.18 Glutamic Acid (Glu, E) -- 5.19 Phenylalanine (Phe, F) -- 5.20 Leucine (Leu, L) -- 5.21 Proline (Pro, P). , 5.22 Serine (Ser, S) -- References -- Chapter 6 Peptide Ligations at Sterically Demanding Sites -- 6.1 Introduction -- 6.2 Ligations Using Thioesters -- 6.2.1 Exogenous Additive‐promoted Ligations -- 6.2.2 Ligations Using Reactive Thioesters -- 6.2.3 Internal Activation Strategy in Peptide Ligations -- 6.3 Ligations Using Oxo‐esters -- 6.4 Peptide Ligations Based on Selenoesters -- 6.5 Microfluidics‐promoted NCL -- 6.6 Representative Applications in Protein Synthesis -- 6.7 Summary and Outlook -- References -- Chapter 7 Controlling Segment Solubility in Large Protein Synthesis -- 7.1 Solvent Manipulation -- 7.2 Isoacyl Strategy -- 7.3 Semipermanent Solubilizing Tags -- 7.3.1 N‐ or C‐Terminal Solubilizing "Tails" -- 7.3.2 Reversible Backbone Modifications as Solubilizing Tags -- 7.3.3 Building Block Solubilizing Tags -- 7.3.4 Extendable Side‐chain‐based Solubilizing Tags -- References -- Chapter 8 Toward HPLC‐free Total Chemical Synthesis of Proteins -- 8.1 Introduction -- 8.1.1 Capture and Release Purification -- 8.1.2 Solid‐phase Chemical Ligations (SPCL) -- 8.2 Synthesis of Peptide Segments for Native Chemical Ligation -- 8.2.1 HPLC‐free Preparation of N‐terminal Peptide Segments for NCL -- 8.2.2 HPLC‐free Preparation of C‐terminal Peptide Segments for NCL -- 8.3 Synthesis of Proteins Using the His6 Tag -- 8.3.1 Reversible His6‐based Capture Tags -- 8.3.2 His6‐based Immobilization for C‐to‐N Assembly of Crambin -- 8.3.3 His6‐based Immobilization for Assembly of Proteins on Microtiter Plates -- 8.3.4 His6 and Hydrazide Tags for Sequential N‐to‐C Capture and Release -- 8.4 Synthesis of Proteins via Oxime Formation -- 8.4.1 Reversible Oxime‐based Capture Tags -- 8.4.2 Oxime‐based Immobilization for N‐to‐C Solid‐phase Chemical Ligations -- 8.4.3 Oxime‐based Immobilization for C‐to‐N Solid‐phase Chemical Ligations. , 8.4.4 Oxime‐based C‐to‐N Solid‐phase Chemical Ligations -- 8.5 Synthesis of Proteins via Hydrazone Formation -- 8.5.1 Reversible Hydrazone‐based Capture Tags -- 8.5.2 Hydrazone‐based Immobilization for Assembly of Proteins on Microtiter Plates -- 8.6 Synthesis of Proteins Using Click Chemistry -- 8.6.1 Click‐based Immobilization for N‐to‐C Solid‐phase Peptide Ligations Using a Protected Alkyne -- 8.6.2 Click‐based Immobilization for N‐to‐C Solid‐phase Peptide Ligations Using a Sea Group -- 8.7 Synthesis of Proteins Using the KAHA Ligation -- 8.7.1 The KAHA Ligation -- 8.7.2 HPLC‐free Synthesis of Proteins Using the KAHA Ligation -- 8.8 Synthesis of Proteins Using Photocleavable Tags -- 8.8.1 Synthesis of Proteins Using a Photocleavable Biotin‐based Purification Tag -- 8.8.2 Synthesis of Proteins Using a Photocleavable His6‐based Purification Tag -- 8.9 Conclusion -- References -- Chapter 9 Solid‐phase Chemical Ligation -- 9.1 Introduction -- 9.1.1 The Promises of Solid Phase Chemical Ligation (SPCL) -- 9.1.2 Chemical Ligation Reactions Used for SPCL -- 9.1.3 Key Requirements for a SPCL Strategy -- 9.2 SPCL in the C‐to‐N Direction -- 9.2.1 Temporary Masking Groups to Enable Iterative Ligations -- 9.2.2 Linkers for C‐to‐N SPCL -- 9.2.2.1 Use of Same Linker and Solid Support for SPPS and SPCL -- 9.2.2.2 Re‐immobilization of the C‐Terminal Segment -- 9.3 SPCL in the N‐to‐C Direction -- 9.3.1 Temporary Masking Groups to Enable Iterative Ligations -- 9.3.2 Linkers for N‐to‐C SPCL -- 9.3.3 Case Study -- 9.3.4 SPCL with Concomitant Purifications -- 9.4 Post‐Ligation Solid‐Supported Transformations -- 9.4.1 Chemical Transformations -- 9.4.2 Biochemical Transformations -- 9.5 Solid Support -- 9.6 Conclusion and Perspectives -- Acknowledgment -- References -- Chapter 10 Ser/Thr Ligation for Protein Chemical Synthesis -- 10.1 Serine/Threonine Ligation. , 10.2 Epimerization Issue.

Note: Intro -- Preface -- Contents -- Chemical Protein Synthesis with the KAHA Ligation -- 1 Introduction -- 1.1 Overview of Chemical Ligations for Protein Synthesis -- 1.2 KAHA Ligation -- 1.2.1 Types of KAHA Ligation -- 1.2.2 Mechanism -- Type I -- Type II -- 2 α-Ketoacids -- 2.1 General Properties of α-Keto Acids -- 2.2 Synthesis of Peptide α-Ketoacids -- 2.2.1 Phosphorus Ylides -- 2.2.2 Sulfur Ylides -- 2.3 Solid Supported Linker for Peptide Sulfur-Ylide Synthesis -- 2.4 Protecting Groups for C-Terminal α-Ketoacids -- 3 Hydroxylamines -- 3.1 Overview -- 3.2 O-Unsubstituted Peptide Hydroxylamines -- 3.2.1 On-Resin Synthesis by Nucleophilic Substitution -- 3.2.2 Nitrone-Protected N-Hydroxy Aminoacid Building Blocks -- 3.2.3 In Situ Preparation of Peptides with N-Terminal Nitrone-Protected N-Hydroxy Aminoacids -- 3.2.4 Synthesis of beta-Peptides with Isoxazolidin-5-One Building Blocks -- 3.3 O-Substituted Hydroxylamines -- 3.3.1 OBz Hydroxylamines -- 3.3.2 O-Aryl Hydroxylamines -- 3.3.3 Cyclic Hydroxylamines -- 4 Ligations/Protein Synthesis -- 4.1 Ligations with Type I Hydroxylamines -- 4.2 Ligations with 5-Oxaproline -- 4.2.1 Formation and Rearrangement of Depsipeptides -- 4.2.2 Two-Segment Ligations with 5-Oxaproline -- 4.2.3 Multi-Segment Ligations -- UFM-1 -- SUMO3 -- 4.3 Peptide Macrocycles -- 4.3.1 Macrocyclizations with Free Hydroxylamines: Synthesis of Epi-Aza-Surfactin -- 4.3.2 Synthesis of Natural Products with Nitrone-Protected Monomers -- 4.3.3 General and Efficient Synthesis of Macrocyclic Peptides with 5-Oxaproline -- 5 Potassium Acyltrifluoroborate (KAT) Ligation -- 6 Outlook -- 6.1 Development of New Cyclic Hydroxylamines -- 6.2 Orthogonally Protected α-Ketoacids -- 6.3 Kinetically Controlled Ligations -- 6.4 Combining KAHA Ligation and Native Chemical Ligation -- 6.5 Summary -- References. , Chemical Synthesis of Proteins Using N-Sulfanylethylanilide Peptides, Based on N-S Acyl Transfer Chemistry -- 1 Introduction -- 2 Development of N-S Acyl Transfer Device for Thioester Synthesis with Practical Applications in Peptide/Protein Synthesis -- 2.1 Naturally Occurring Thioester Formation: Intein-Extein System -- 2.2 Chemistry Seen in Naturally Occurring Thioester Formation -- 2.3 Peptidyloxazolidinone as Thioester Precursor -- 2.4 N-Sulfanylethylaniline Linker as Alternative to Oxazolidinone -- 2.5 Initial Observation on of SEAlide Peptide -- 2.6 Attempted Sequential NCL Using SEAlide Peptides -- 2.7 SEAlide Peptides Function as Thioesters in the Presence of Phosphate Salts -- 2.8 Synthesis of hANP by One-Pot/N-to-C-Directed Sequential NCL -- 2.9 One-Pot/Four-Segment Ligation -- 2.10 Dual Kinetically Controlled Ligation -- 2.11 Chemical Synthesis of Proteins Using SEAlide Peptides -- 3 Summary, Conclusions, and Outlook -- References -- Postligation-Desulfurization: A General Approach for Chemical Protein Synthesis -- 1 Introduction -- 2 Postligation-Desulfurization (Alanine Ligation) -- 2.1 Metal-Based Desulfurization -- 2.2 Metal-Free Desulfurization -- 2.3 Application of Postligation-Metal-Free Desulfurization (Ala Ligation) -- 3 Postligation-Desulfurization at Other Amino Acid Sites -- 3.1 Metal-Free Desulfurization -- 3.1.1 Ligation at Valine Site -- 3.1.2 Ligation at Lysine Site -- 3.1.3 Ligation at Threonine Site -- 3.1.4 Ligation at Leucine Site -- 3.1.5 Ligation at Proline Site -- 3.1.6 Ligation at Arginine Site -- 3.1.7 Ligation at Aspartic Acid Site -- 3.1.8 Ligation at the Glutamate Site -- 3.2 Metal-Based Desulfurization -- 3.2.1 Ligation at the Phenylalanine Site -- 3.2.2 Ligation at the Glutamine Site -- 3.2.3 Ligation at the Tryptophan Site -- 3.3 Sugar-Assisted Ligation (SAL) -- 3.4 One-Pot Ligation-Desulfurization. , 4 Total Synthesis of hPTH -- 5 Conclusion -- References -- Solid Phase Protein Chemical Synthesis -- 1 Introduction -- 2 Chemical Protein Synthesis by Stepwise Solid Phase Peptide Synthesis -- 3 Purification by Selective Capture on a Solid Support -- 3.1 Purification by Covalent Capture on a Solid Support -- 3.2 Purification by Covalent Internal Resin Capture -- 3.3 Purification by Selective and Non-Covalent Adsorption on a Solid Support -- 4 Chemical Protein Synthesis by the Solid Phase Sequential Chemoselective Ligation of Unprotected Peptide Segments -- 4.1 Advantages of the Solid Phase Approach -- 4.2 Solid Phase Protein Synthesis in the C-to-N Direction -- 4.2.1 N-Terminal Cysteine Protection Strategies -- 4.2.2 Linker Strategies for C-to-N Solid Phase Elongation Strategies -- 4.3 N-to-C Solid Phase Chemical Protein Synthesis -- 5 Conclusion -- References -- New Methods for Chemical Protein Synthesis -- 1 Introduction -- 2 Synthesis of Peptide Fragments -- 2.1 New Methods for Peptide Synthesis -- 2.1.1 Amide Bond Formation -- 2.1.2 Methods for the Synthesis of Long/Difficult Peptides -- 2.2 New Methods for Activated Peptide Synthesis -- 3 Assembly of Peptide Fragments -- 3.1 New Methods for Peptide Ligation -- 3.1.1 KAHA Method -- 3.1.2 Fragment Condensation -- 3.1.3 Ser and Thr Ligation -- 3.2 One-Pot Strategies for the Assembly of Peptide Fragments -- 3.2.1 C-to-N Direction -- 3.2.2 N-to-C Direction -- 4 Folding of Synthetic Polypeptide Chains -- 5 Conclusion -- References -- Chemical and Biological Tools for the Preparation of Modified Histone Proteins -- 1 Introduction -- 2 Genetic Approaches for Modified Histones -- 2.1 Genetic Mimics of Histone Modifications -- 2.2 Codon Suppression: Expanded Genetic Code Approaches to Modified Histones -- 2.2.1 Encoded Lysine Modifications. , 2.2.2 Combined Genetic and Chemical Approaches: Modifications Introduced Through Dehydroalanine -- 2.2.3 Encoded Phosphoserine -- 3 Chemical Installation of PTM Analogs at Single Cysteine Sites -- 3.1 MLAs: Methyllysine Analogs -- 3.2 Acetyllysine Analogs via Cysteine Alkylation -- 3.3 Thiol-ene Chemistry to Introduce Modification Analogs -- 3.4 Disulfide Stapling -- 4 Chemical Ligation for the Preparation of Modified Histone Proteins -- 4.1 Histone Semi-Synthesis by Expressed Protein Ligation: Modifications Near the Histone N-Terminus -- 4.2 Considerations for Selection of Appropriate Ligation Sites -- 4.3 Histone Semi-Synthesis by Expressed Protein Ligation: Modifications Near the Histone C-Terminus -- 4.4 Total Synthesis of Histone Proteins by NCL -- 5 Prospects: Synthetic Histone Proteins in the Eukaryotic Cell -- References -- Index.

Note: Intro -- Preface -- Contents -- Total Synthesis of Glycosylated Proteins -- 1 Introduction -- 2 Chemical Synthesis of Glycopeptides -- 3 Chemical Synthesis of Glycoproteins -- 3.1 Advances in Chemical Ligation -- 3.2 Total Synthesis of Homogeneous N-Glycosylated Proteins -- 3.2.1 Human Glycoprotein Hormones -- Human α-Glycoprotein Hormone (α-hGPH) -- Human beta-Follicle Stimulating Hormone (beta-hFSH) -- Human beta-Luteinizing Hormone (beta-hLH) and beta-ChorionicGonadotropin Hormone (beta-hCG) -- 3.2.2 Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF) -- 3.2.3 Erythropoietin (EPO) -- 4 Conclusion -- References -- Modern Extensions of Native Chemical Ligation for Chemical Protein Synthesis -- 1 Introduction -- 2 Native Chemical Ligation -- 2.1 Scope and Mechanism -- 2.2 Modern Application -- 3 Development of New Cysteine Ligation Surrogates -- 3.1 Auxiliary-Based Methods -- 3.1.1 N-Terminal Ligation Auxiliaries -- 3.1.2 Side-Chain Ligation Auxiliaries -- 4 Post-Ligation Manipulations -- 4.1 Ligation-Desulfurization -- 4.2 Ligation-Desulfurization in Protein Synthesis -- 4.3 Ligation-Desulfurization at Thiol-Derived Amino Acids -- 4.3.1 Phenylalanine -- 4.3.2 Valine -- 4.3.3 Threonine -- 4.3.4 Lysine -- 4.3.5 Leucine -- 4.3.6 Proline -- 4.3.7 Glutamine -- 4.3.8 Arginine -- 4.3.9 Aspartic Acid -- 4.3.10 Glutamic Acid -- 4.3.11 Tryptophan -- 4.3.12 Protein Synthesis via Ligation at Non-Cys Sites -- 4.4 Ligation at Selenocysteine -- 4.5 Ligation-Deselenization Chemistry -- 5 Conclusion -- References -- Chemical Methods for Protein Ubiquitination -- 1 Introduction -- 2 Preparation of Ub-Proteins with Native Isopeptide Bonds -- 2.1 Nα-Auxiliary-Mediated Ubiquitination -- 2.2 Thiolated-Lysine-Mediated Ubiquitination -- 2.3 Ubiquitination Through Ag+-Mediated Activation of Ubiquitin C-Terminal Thioester. , 3 Preparation of Ub-Proteins with Non-Native Linkages -- 4 Conclusion -- References -- Peptide Thioester Formation via an Intramolecular N to S Acyl Shift for Peptide Ligation -- 1 Introduction -- 2 Intramolecular N to S Acyl Shift Reaction -- 2.1 At the Thiol Auxiliary, Dmmb -- 2.2 At the Cysteine Residue -- 3 Peptide Thioester Synthesis via Intramolecular N to S Acyl Shift -- 3.1 Active Acyl Structure -- 3.2 Thiol Auxiliary, Dmmb -- 3.3 tert-Amide with Vicinal Thiol Group -- 3.4 tert-Amide with Irreversibility -- 3.5 C-Terminal Cysteine -- 3.6 Cysteinylproline Ester (CPE) -- 3.7 Cysteinylprolylcysteine (CPC) -- 4 Protein Synthesis via N to S Acyl Shift Based on the CPE System -- Conclusions -- References -- Chemical Synthesis and Biological Function of Lipidated Proteins -- 1 Introduction -- 1.1 N-Myristoylation -- 1.2 Palmitoylation -- 1.3 Prenylation -- 1.4 GPI-Anchor Addition -- 1.5 Other Types of Lipidation -- 2 Synthesis of Lipidated Peptides -- 2.1 Preparation of Lipidated Cysteine Building Blocks -- 2.2 Solution-Phase Synthesis of Lipidated Peptides -- 2.3 Solid-Phase Approach for the Synthesis of Lipidated Peptides -- 2.4 Synthesis of Lipidated Peptides by Combined Solution/Solid-Phase Approach -- 2.4.1 Synthesis of Phosphatidylethanolaminylated Peptide -- 2.4.2 Synthesis of Sterol-Modified Peptide -- 3 Synthesis of Lipidated Proteins -- 3.1 Assisted Solubilisation Strategy -- 3.2 Expressed Protein Ligation -- 3.3 MIC Ligation -- 3.4 Diels-Alder Ligation -- 3.5 Click Ligation -- 3.6 Sortase-Mediated Protein Ligation -- 4 Chemical Biology of Lipidated Protein -- 4.1 Cell Biological Studies of S-Palmitoylation Cycle of Ras GTPases -- 4.2 Biophysical Studies of Lipidated Ras GTPases -- 4.3 Structural Studies of Prenylated Rheb GTPases -- 4.4 Thermodynamic Basis of Rab GTPases Membrane Targeting -- 4.5 Biological Function of GPI-Anchors. , 4.6 Function of LC3-PE in Autophagosome Formation -- 5 Conclusions and Perspectives -- References -- Protein Chemical Synthesis in Drug Discovery -- 1 Introduction -- 2 A Brief History of Modern Chemical Protein Synthesis -- 2.1 Peptide Chemistry in the Early Years, 1880s-1950s -- 2.2 Solid-Phase Peptide Synthesis (SPPS), 1960s-1980s -- 2.3 Protein Synthesis by Peptide Ligation, 1990s-Present -- 2.4 Analytical Methods -- 3 Key Chemistries for Modern Chemical Protein Synthesis -- 3.1 Native Chemical Ligation -- 3.2 Preparation of Peptide C-Terminal Thioesters -- 3.2.1 Preparation of Peptide C-Terminal Thioester by Boc-SPPS -- 3.2.2 Preparation of Peptide C-Terminal Thioesters by Fmoc-SPPS -- 3.2.3 Preparation of Protein C-Terminal Thioesters by Expressed Protein Ligation -- 3.3 Ligation at Non-Cys Junctions -- 3.3.1 Thiol Handle-Mediated Ligation at Non-Cys Junction -- 3.3.2 Thiol Handle-Free Ligation at Non-Cys Junction -- 3.4 Multiple Segment Ligation Strategies -- 3.5 The Folding of a Synthetic Protein -- 4 Applications of Protein Chemical Synthesis to Drug Discovery -- 4.1 Chemical Synthesis of d-Proteins and Their Application to Drug Discovery -- 4.1.1 Mirror Image Phage-Display -- 4.1.2 Racemic Protein Crystallization -- 4.2 Chemical Synthesis of Enzymes -- 4.3 Protein Pharmaceuticals -- 4.3.1 Chemical Synthesis of Erythropoietin -- 4.3.2 Chemical Synthesis of Insulin -- 5 Conclusion -- References -- Applications of Chemical Ligation in Peptide Synthesis via Acyl Transfer -- 1 Introduction to Native Chemical Ligation -- 2 Solid Phase vs Solution Phase Chemical Ligation -- 3 Intramolecular Chemical Ligation (Acyl Migration) -- 3.1 S- to N-Acyl Migration -- 3.2 O- to N-Acyl Migrations -- 3.3 N- to N-Acyl Migrations -- 4 Applications of Native Chemical Ligation -- 4.1 Synthesis of Cyclic Peptides -- 4.2 Synthesis of Glycopeptides. , 5 Computational Rationalization of Chemical Ligation -- 6 Conclusions -- References -- Index.