Perkins Lab
Single Molecule Biophysics

MCDB 4550/5550 Syllabus: Cellular and Molecular Motion, A Biophysical Approach

Instructor: Thomas Perkins, tperkins@jila.colorado.edu 303-492-5291

This course provides an overview of the biophysics of enzymes, nucleic acids and the cytoskeleton. Straightforward application of classical Newtonian physics does a poor job of providing insight into biological processes. This course will develop a simple description of the biophysics governing the motion of molecules inside of cells. Topics covered will include diffusion, molecular motors, polymerization of the cytoskeleton and the polymer properties of nucleic acids and microtubules. Each student will give two oral presentations on journal articles along with a critique of the research article. Problem sets will provide a quantitative understanding of the topics discussed. A final paper will be required that addresses a critical question in biophysics and/or outlines a proposed experimental approach to answer the question.

Prereq., CHEM 1131 or 1171, physics 2010, 2020, MCDB 3120, or instructor consent. Recommended prereq., Math 1300 and/or CHEM 3311. Same as MCDB 5550.

WeekTuesdayThursday
1: 1/17-19 Introduction Reading: Chapter 1 Random walks & mechanical forces Reading: Chapter 2
Problem Set #1
2: 1/24-26 Mass, stiffness & damping of a protein Reading: Chapter 3 Purcell: Life at low Reynolds number
Problem Set #2
3: 1/31-2/2 Thermal forces & diffusion Reading: Chapter 4 Cluzel et al., An Ultrasensitive bacterial motor
Problem Set #3
4: 2/7-9 Chemical forces Reading: Chapter 5 Wang et al., Force and Velocity Measured for single RNAP
Problem Set #4
5: 2/14-16 Polymer mechanics Reading: Chapter 6 Wuite et al., Single molecule studies of T7 DNA polymerase
6: 2/21-23 No Class Structure and Mechanics of Cytoskeletal: Reading: Chapter 7 & 8
7: 2/28-3/2 Polymerization of filaments Reading: Chapter 9 Waterman-Storer et al., New Directions in Fluorescent Speckle Microscopy
Problem Set #5
8: 3/7-9 Force generation by cytoskeletal filaments Reading: Chapter 10 Dogterom et al, Force-Velocity relation for growing microtubules
Problem Set #6
9: 3/14-16 Active Polymerization Reading: Chapter 11 Cameron et al., Motility of ActA protein-coated beads driven by Actin
Problem Set #7
10: 3/21-23 Structure and Speed of motor proteins Reading: Chapter 12 & 13 Kural et al., Kinesin and Dynein move a peroxisome in Vivo
Problem Set #8
11: 4/4-6 ATP Hydrolysis Reading: Chapter 14 Adachi et al., Stepping of F1ATPase resolved with single fluorophores
Problem Set #9
12: 4/11-13 Steps and forces Reading: Chapter 15 Visscher et al., Single kinesin molecules studies with a force clamp
Problem Set #10
13: 4/18-20 Motility models Reading: Chapter 16 Ruff et al., Single-molecule tracking with genetically engineered myosin
Problem Set #11
14: 4/25-27 To be announced. Hua et al., Kinesin neck does not rotate
Problem Set #12
15: 5/2-4 Guest Lecture: Dick McIntosh Summary
Final paper due May 5th (click for details)

Critical Review

Each student will write a 2 page critique on the papers that they present orally to the class (due the second class after their lecture). This summary will be graded and distributed to the class. Prior to a class covering a paper, students will be given a series of questions that can be answered by reading the paper. Students will also provide set of at least three well thought out questions for the student presenter. Students will prepare a final written paper that addresses a critical question in biophysics and outlines a proposed experimental approach to answer the question.

Paper

Each student will write a 6-8 page paper (10-12 for graduate students) that either proposes (i) an experiment using biophysical techniques with quantitative estimates, (ii) a critique of a set of papers, or (iii) a topic mutually agreed upon between the student and the instructor.

Presentations

Each student will give two oral presentations, in teams if necessary, on the primary literature reading assignments. These presentations will be followed by a discussion led by the presenting student(s) to critically analyze the reading material and to review what students have learned from the presentation and the reading material.

Grade determination

20% Weekly critical writings 20% Participation in class discussion 20% Oral presentation 20 % Problem sets 20 % Final written proposal

Texts (required for course)

Mechanics of Motor Proteins and the Cytoskeleton By Jonathon Howard, Sinauer, 2001

Texts (supplementary information)

Molecular Biology of the Cell (4th Edition, silver cover), By Alberts, et al., 2002 A good, encyclopedic biology text book. Very good for learning vocabulary but a little dry

Biochemistry (5th Edition, green cover), By Berg, Stryer, & Tymoczko, 2002 A great book for learning about biochemistry. Clearly explains lots of topics but not as extensive MBOC. My first choice when I am learning about biochemistry. Older versions of either of these books would be sufficient for this class.

Office Hours

Office: JILA A503 Office Hours: Wednesday 2-3 pm