Kinetics of DNA translocase SpoIIIE studied by dual optical tweezers

Lin Ling; E Qu; Honglian Guo; Zhaolin Li; Lu Huang; Daozhong Zhang; Zhiyuan Li

doi:10.1117/12.837761

1 December 2009 Kinetics of DNA translocase SpoIIIE studied by dual optical tweezers

Lin Ling, E Qu, Honglian Guo, Zhaolin Li, Lu Huang, Daozhong Zhang, Zhiyuan Li

Proceedings Volume 7507, 2009 International Conference on Optical Instruments and Technology: Optical Trapping and Microscopic Imaging; 750707 (2009) https://doi.org/10.1117/12.837761
Event: International Conference on Optical Instrumentation and Technology, 2009, Shanghai, China

Abstract

DNA translocase SpoIIIE protein is a kind of motor proteins, which transports DNA from one side of the membrane to the other side, so it plays an important role in cell division. In experiment, λDNA is labeled on one end with biotin and the other with digoxigenin. In this work we study kinetics of DNA translocase SpoIIIE by means of dual optical tweezers. In our experiment, λDNA is tethered between streptavidin-coated polystyrene bead and antidigoxigenin-coated polystyrene bead held by dual optical tweezers. One trap is immovable, and the other is movable. When SpoIIIE protein transports DNA, the length of DNA changes. The length change can be calculated according to the displacement of the trapped bead, which is detected by quadrant photodiode. When SpoIIIE transports DNA, DNA is shortened by up to about 500nm, then as the translocation stops, the DNA returns to its normal length, and this process repeats time and time again. The most probable speed that SpoIIIE transports DNA is 710nm/s.

Citation Download Citation

Lin Ling, E Qu, Honglian Guo, Zhaolin Li, Lu Huang, Daozhong Zhang, and Zhiyuan Li "Kinetics of DNA translocase SpoIIIE studied by dual optical tweezers", Proc. SPIE 7507, 2009 International Conference on Optical Instruments and Technology: Optical Trapping and Microscopic Imaging, 750707 (1 December 2009); https://doi.org/10.1117/12.837761

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