INVESTIGATION OF DYNEIN’S
LINKER REGIONS
Mentor: Dr. Elisar Barbar
Jonathan Yih
RELEVANCE
 Role
in basic life
processes
http://hopes.stanford.edu/basics/dna/f_b14mit_meta.jpg
 Insight
for dyneinassociated problems
http://www.thehealthsuccesssite.com/images/muscle-als.jpg
GOAL
 Improve
understanding of dynein’s structure
http://www.physics.berkeley.edu/research/yildiz/DyneinNew.jpg
BACKGROUND
 Motor

protein
Energy
Work
 Functions:

Transport

Orienting &
positioning organelles

Cell division
http://maepro.uta.edu/loco/pictures/kinesin11.jpg
BACKGROUND
 Dynein’s
structure
Heavy Heads
and Chains
Cargo
Microtubule
IC
LC8
• Light & Intermediate Chains • Heavy Heads & Chains
Binding
ATP hydrolysis & movement
HYPOTHESIS
 LC8
binding to IC initiates the
assembly of the dynein complex.
OVERVIEW
 Surface
Plasmon Resonance

Study protein kinetics

LC8 with:
 IC

nNOS

BIM
LC8/IC compex image © Barbar 2007

Swa
SURFACE PLASMON RESONANCE
Reflected Light
Incident Light
Analyte
Immobilized
Protein (LC8)
Gold Surface
Results!
SURFACE PLASMON RESONANCE
Protein complex
formation
A+B
ka
kd
http://www.discoversensiq.com/uploads/file/support/spr/Overview_of_SPR.pdf
AB
SURFACE PLASMON RESONANCE
 Immobilization
3
2
1
Time (s)
SPR: LC8 & BIM OR NNOS
SPR: LC8 & IC
Res SD 6.704
SPR: EXPERIMENTAL DESIGN
 Replications
 Multiple
concentrations (5+)
 Adequate
 Reduce
regeneration
mass transport limitations
 Achieve
steady state
ACKNOWLEDGEMENTS
 Special


Dr. Elisar Barbar
The Barbar Lab Group




thanks to:
Afua Nyarko,
Yujuan Song, & Jessica Morgan
Nathan Gillock & Aaron Martin
Dr. Kevin Ahern
The Howard Hughes Medical Institute