i nvestigation of d ynein ’ s l inker r egions mentor: dr. elisar barbar jonathan yih
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INVESTIGATION OF DYNEIN’S LINKER REGIONSMentor: Dr. Elisar Barbar
Jonathan Yih
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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
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GOAL
Improve understanding of dynein’s structure
http://www.physics.berkeley.edu/research/yildiz/DyneinNew.jpg
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BACKGROUND
Motor protein
Energy Work
Functions:
Transport
Orienting & positioning organelles
Cell division http://maepro.uta.edu/loco/pictures/kinesin11.jpg
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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
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HYPOTHESIS
LC8 binding to IC initiates the assembly of the dynein complex.
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OVERVIEW
Surface Plasmon ResonanceStudy protein kinetics
LC8 with:IC
nNOS
BIM
Swa
LC8/IC compex image © Barbar 2007
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SURFACE PLASMON RESONANCE
Gold Surface
Results!
Immobilized Protein (LC8)
Incident Light Reflected Light
Analyte
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SURFACE PLASMON RESONANCE
http://www.discoversensiq.com/uploads/file/support/spr/Overview_of_SPR.pdf
Protein complex formation
A + B ABka
kd
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SURFACE PLASMON RESONANCE
Immobilization
1
2
3
Time (s)
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SPR: LC8 & BIM OR NNOS
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SPR: LC8 & IC
Res SD 6.704
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SPR: EXPERIMENTAL DESIGN
Replications
Multiple concentrations (5+)
Adequate regeneration
Reduce mass transport limitations
Achieve steady state
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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