Kinetic Analysis of the Interaction of Mos1 Transposase with its Inverted Terminal Repeats Reveals New Insight into the Protein–DNA Complex Assembly

摘要: Transposases are specific DNA-binding proteins that promote the mobility of discrete DNA segments. We used a combination physicochemical approaches to describe association MOS1 (an eukaryotic transposase) with its target DNA, an event corresponding first steps transposition cycle. Because kinetic constants reaction still unknown, we aimed determine them by using quartz crystal microbalance on two sources recombinant MOS1: one produced in insect cells and other bacteria. The prokaryotic-expressed showed no cooperativity displayed Kd about 300 nM. In contrast, eukaryotic-expressed generated cooperative system, lower (∼2 nm). origins these differences were investigated IR spectroscopy AFM imaging. Both support conclusion prokaryotic- not similarly folded, thereby resulting early transposition.