This second edition volume expands on the previous edition with a discussion of the latest experimental and computational strategies used in the study, characterization, and targeting of enzymes with therapeutic potential. The chapters in the book are organized into two parts: Part One presents detailed, reproducible protocols for enzyme characterization, inhibitor screening, and biochemical analysis. Part Two is dedicated to computational and in silico methodologies, reflecting the growing impact of computational biology in drug discovery. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and authoritative, Targeting Enzymes for Pharmaceutical Development: Methods and Protocols, Second Edition is a valuable resource for academic and industrial scientists in medicinal chemistry, chemical biology, pharmaceutical biotechnology, structural biology, and computational drug discovery. This book is both a technical manual and conceptual guide for scientists engaged in enzyme-related drug discovery and development.