INVESTIGATION OF DYNEIN’S LINKER REGIONSMentor: Dr. Elisar Barbar

Jonathan Yih

RELEVANCE

Role in basic life processes

http://hopes.stanford.edu/basics/dna/f_b14mit_meta.jpg

Insight for dynein-associated 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 & Chains

ATP hydrolysis & movement

• Light & Intermediate Chains

Binding

IC

LC8

Cargo

Heavy Heads and Chains

Mic

rotu

bu

le

HYPOTHESIS

LC8 binding to IC initiates the assembly of the dynein complex.

OVERVIEW

Surface Plasmon ResonanceStudy protein kinetics

LC8 with:IC

nNOS

BIM

Swa

LC8/IC compex image © Barbar 2007

SURFACE PLASMON RESONANCE

Gold Surface

Results!

Immobilized Protein (LC8)

Incident Light Reflected Light

Analyte

SURFACE PLASMON RESONANCE

http://www.discoversensiq.com/uploads/file/support/spr/Overview_of_SPR.pdf

Protein complex formation

A + B ABka

kd

SURFACE PLASMON RESONANCE

Immobilization

1

2

3

Time (s)

SPR: LC8 & BIM OR NNOS

SPR: LC8 & IC

Res SD 6.704

SPR: EXPERIMENTAL DESIGN

Replications

Multiple concentrations (5+)

Adequate regeneration

Reduce mass transport limitations

Achieve steady state

ACKNOWLEDGEMENTS

Special thanks to:Dr. Elisar BarbarThe Barbar Lab Group

Afua Nyarko, Yujuan Song, & Jessica Morgan

Nathan Gillock & Aaron MartinDr. Kevin AhernThe Howard Hughes Medical Institute