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DNA Repair Enzymes, Part B, Volume 592 is the latest volume in the Methods in Enzymology series and the first part of a thematic that focuses on DNA Repair Enzymes. Topics in this updated volume include MacroBac: New Technologies for Robust and Efficient Large-Scale Production of Recombinant Multiprotein Complexes, Production and Assay of Recombinant Multisubunit Chromatin Remodeling Complexes, Analysis of Functional Dynamics of Modular Multidomain Proteins by SAXS and NMR, the Use of Single-Cysteine Variants for Trapping Transient States in DNA Mismatch Repair, and Structural Studies of RNases H2 as an Example of Crystal Structure Determination of Protein-Nucleic Acid Complexes. - Includes contributions from leading authorities working in enzymology - Focuses on DNA repair enzymes - Informs and updates on all the latest developments in the field of enzymology
This volume is a timely and comprehensive description of the many facets of DNA and RNA modification-editing processes and to some extent repair mechanisms. Each chapter offers fundamental principles as well as up to date information on recent advances in the field (up to end 2008). They ended by a shortconclusion and future prospect' section and
Since the discovery of the first examples of 2-oxoglutarate-dependent oxygenase-catalysed reactions in the 1960s, a remarkably broad diversity of alternate reactions and substrates has been revealed, and extensive advances have been achieved in our understanding of the structures and catalytic mechanisms. These enzymes are important agrochemical targets and are being pursued as therapeutic targets for a wide range of diseases including cancer and anemia. This book provides a central source of information that summarizes the key features of the essential group of 2-oxoglutarate-dependent dioxygenases and related enzymes. Given the numerous recent advances and biomedical interest in the field, this book aims to unite the latest research for those already working in the field as well as to provide an introduction for those newly approaching the topic, and for those interested in translating the basic science into medicinal and agricultural benefits. The book begins with four broad chapters that highlight critical aspects, including an overview of possible catalytic reactions, structures and mechanisms. The following seventeen chapters focus on carefully selected topics, each written by leading experts in the area. Readers will find explanations of rapidly evolving research, from the chemistry of isopenicillin N synthase to the oxidation mechanism of 5-methylcytosine in DNA by ten-eleven-translocase oxygenases.
The book is concerned with features analysis of radiation damage to living matter at different levels of its organisation, beginning from the molecular level. The general theory of the mechanisms of biological action of radiation based on four logically connected principles of radiation biology is presented for the first time. The book contains abundant material on primary and secondary processes of the response of living systems to radiation, the mechanisms of direct and indirect action of radiation through radiolysis of water and lipids; a detailed consideration of the theory of targets (DNA and biological membranes) and the problems of high- and low-dose irradiation in the context of present-day. The systemic response of the cell to irradiation (oxidative stress, endogenous background radioresistance, damage and repair of biological membranes and DNA, systemic protection against radiation) is approximated. Presentation of these important problems is preceded by an up-to-date evaluation of some aspects regarding dosimetry of ionising radiation.
DNA Repair Enzymes, Part A, Volume 591 is the latest volume in the Methods in Enzymology series and the first part of a thematic that focuses on DNA repair enzymes. Topics in this new release include chapters on the Optimization of Native and Formaldehyde iPOND Techniques for Use in Suspension Cells, the Proteomic Analyses of the Eukaryotic Replication Machinery, DNA Fiber Analysis: Mind the Gap!, Comet-FISH for Ultrasensitive Strand-Specific Detection of DNA Damage in Single Cells, Examining DNA Double-Strand Break Repair in a Cell Cycle-Dependent Manner, Base Excision Repair Variants in Cancer, and Fluorescence-Based Reporters for Detection of Mutagenesis in E. coli. - Includes contributions from leading authorities working in enzymology - Focuses on DNA repair enzymes - Informs and updates on all the latest developments in the field of enzymology
Single-molecule techniques eliminate ensemble averaging, thus revealing transient or rare species in heterogeneous systems [1–3]. These approaches have been employed to probe myriad biological phenomena, including protein and RNA folding [4–6], enzyme kinetics [7, 8], and even protein biosynthesis [1, 9, 10]. In particular, immobilization-based fluorescence te- niques such as total internal reflection fluorescence microscopy (TIRF-M) have recently allowed for the observation of multiple events on the millis- onds to seconds timescale [11–13]. Single-molecule fluorescence methods are challenged by the instability of single fluorophores. The organic fluorophores commonly employed in single-molecule studies of biological systems display fast photobleaching, intensity fluctuations on the millisecond timescale (blinking), or both. These phenomena limit observation time and complicate the interpretation of fl- rescence fluctuations [14, 15]. Molecular oxygen (O) modulates dye stability. Triplet O efficiently 2 2 quenches dye triplet states responsible for blinking. This results in the for- tion of singlet oxygen [16–18]. Singlet O reacts efficiently with organic dyes, 2 amino acids, and nucleobases [19, 20]. Oxidized dyes are no longer fluor- cent; oxidative damage impairs the folding and function of biomolecules. In the presence of saturating dissolved O , blinking of fluorescent dyes is sup- 2 pressed, but oxidative damage to dyes and biomolecules is rapid. Enzymatic O -scavenging systems are commonly employed to ameliorate dye instability. 2 Small molecules are often employed to suppress blinking at low O levels.