Using a Nanopore to Watch Enzymes at Work
Event details
| Date | 05.11.2018 |
| Hour | 12:15 |
| Speaker | Prof. Jens H. Gundlach, Department of Physics, University of Washington, Seattle, Washington (USA) |
| Location | |
| Category | Conferences - Seminars |
DISTINGUISHED LECTURE IN BIOLOGICAL ENGINEERING
(sandwiches served)
Abstract:
We have engineered a high-resolution molecular sensor based on an ion current flowing through the protein nanopore MspA. This sensor has enabled nanopore sequencing of DNA. Here, I will show how we have further adapted this nanopore technology as a single-molecule tool called SPRNT (Single molecule Picometer Resolution Nanopore Tweezers). SPRNT permits real-time observation of enzyme mechanics with unprecedented spatio-temporal resolution while it simultaneously reveals the nucleotide sequence within the enzyme. I will show hereto unseen detail in the motion of several different enzymes along nucleic acids. In the helicase Hel308, we observe two steps, an ATP-dependent and an ATP-independent step, for each full nucleotide progression by the enzyme. The stepping kinetics depends on the sequence of the DNA substrate. In addition, using the core polymerase of the transcription complex (RNAP), we observe detailed enzyme activity at sequence-specific pausing sites.
Bio:
Physik-Vordiplom , Johannes Gutenberg Universit¨at, Mainz, Germany, 1982.
Physik-Diplom , Johannes Gutenberg Universit¨at, Mainz, Germany, 1986.
Ph.D. (Nuclear Physics), University of Washington, 1990.
Postdoctoral Research Associate (Experimental Gravitation),
University of Washington, 1990-93.
Research Assistant Professor, University of Washington, 1993-98.
Research Associate Professor, University of Washington, 1998-2004.
Full Professor, University of Washington, 2004-present.
(sandwiches served)
Abstract:
We have engineered a high-resolution molecular sensor based on an ion current flowing through the protein nanopore MspA. This sensor has enabled nanopore sequencing of DNA. Here, I will show how we have further adapted this nanopore technology as a single-molecule tool called SPRNT (Single molecule Picometer Resolution Nanopore Tweezers). SPRNT permits real-time observation of enzyme mechanics with unprecedented spatio-temporal resolution while it simultaneously reveals the nucleotide sequence within the enzyme. I will show hereto unseen detail in the motion of several different enzymes along nucleic acids. In the helicase Hel308, we observe two steps, an ATP-dependent and an ATP-independent step, for each full nucleotide progression by the enzyme. The stepping kinetics depends on the sequence of the DNA substrate. In addition, using the core polymerase of the transcription complex (RNAP), we observe detailed enzyme activity at sequence-specific pausing sites.
Bio:
Physik-Vordiplom , Johannes Gutenberg Universit¨at, Mainz, Germany, 1982.
Physik-Diplom , Johannes Gutenberg Universit¨at, Mainz, Germany, 1986.
Ph.D. (Nuclear Physics), University of Washington, 1990.
Postdoctoral Research Associate (Experimental Gravitation),
University of Washington, 1990-93.
Research Assistant Professor, University of Washington, 1993-98.
Research Associate Professor, University of Washington, 1998-2004.
Full Professor, University of Washington, 2004-present.
Practical information
- Informed public
- Free
Organizer
Contact
- Institute of Bioengineering (IBI, Christina Mattsson)