Single-molecule studies of complex systems: the replisome.
A complete, system-level understanding of biological processes requires comprehensive information on the kinetics and thermodynamics of the underlying biochemical reactions. A wide variety of structural, biochemical, and molecular biological techniques have led to a quantitative understanding of the molecular properties and mechanisms essential to the processes of life. Yet, the ensemble averaging inherent to these techniques limits us in understanding the dynamic behavior of the molecular participants. Recent advances in imaging and molecular manipulation techniques have made it possible to observe the activity of individual enzymes and record "molecular movies" that provide insight into their dynamics and reaction mechanisms. An important future goal is extending the applicability of single-molecule techniques to the study of larger, more complex multi-protein systems. In this review, the DNA replication machinery will be used as an example to illustrate recent progress in the development of various single-molecule techniques and its contribution to our understanding of the orchestration of multiple enzymatic processes in large biomolecular systems.
van Oijen
Antoine M
AM
Harvard Medical School, Dept. of Biological Chemistry and Molecular Pharmacology, 240 Longwood Avenue, Boston, MA 02115, USA. antoine_van_oijen@hms.harvard.edu
D000818:Animals; D004265:DNA Helicases / Q000502:physiology; D004261:DNA Replication / Q000502:physiology*; D004259:DNA-Directed DNA Polymerase / Q000502:physiology; D006801:Humans; D008954:Models, Biological; D015336:Molecular Probe Techniques; D009097:Multienzyme Complexes / Q000502:physiology*; D035602:Transcriptional Elongation Factors / Q000502:physiology
D009097:Multienzyme Complexes; D035602:Transcriptional Elongation Factors; D004259:DNA-Directed DNA Polymerase; D004265:DNA Helicases