· Translate this pagePK ! ¦nFi N [Content_Types].xml ¢ (
Transcript of · Translate this pagePK ! ¦nFi N [Content_Types].xml ¢ (
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kScience!Comparing GEF-induced GTP binding between the
Hlavacek and Faeder models, using Stites 2007 paper rates
Eitan Pearl, Stephanie Salarbux, Jennifer Nam
CMACS Workshop 2014
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The Stites Paper Minimal Model(no intermediate apo-Ras steps)
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Inputting the Stites differential equation for the GEF-nt-exchange rate
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The Hlavacek Model: Allosteric Binding Reactions
We added the rates for positive feedback loop – allosteric SOS1 binding (from Cell 2009)Ras
Unbound, slow rate Ras-GDP
Bound, intermediate rate, 5x faster Ras-GTP
Bound, Max allosteric rate, 75x faster
Then compared the Hlavacek (Mass-Action) and Faeder (Minimal) Models
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Hlavacek, Faeder Models with Stites’ Rates: Conversions Required!
Stites paper rates into Hlavacek model:
Increased SOS1 concentrations Parameter scan and gdat
simulations
Stites paper rates into Faeder model:
Added units to be comparable to the Hlavacek model
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Hlavacek Model
Increased SOS1 binding rates in the model to Stites paper rates/concentrationsEffect: turned off Ras-GDP Ras-GTP conversion
Possible Mutation: Turned off GAP (GTP hydrolysis)
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Allosteric Binding in the Hlavacek Model:Changing Rate of Fast SOS1
0.00E+00 5.00E+02 1.00E+03 1.50E+03 2.00E+03 2.50E+03 3.00E+03 3.50E+030.00E+00
5.00E+04
1.00E+05
1.50E+05
2.00E+05
2.50E+05
RasGTPRasGDP
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Allosteric Binding in the Hlavacek Model:Changing Rate of Basic SOS1
0.00E+00 5.00E+02 1.00E+03 1.50E+03 2.00E+03 2.50E+03 3.00E+03 3.50E+030.00E+00
5.00E+04
1.00E+05
1.50E+05
2.00E+05
2.50E+05
RasGTPRasGDP
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What happens when you increase Sos?Simulation
Gdat shows crossover of GTP and GDPParameter Scan
Interpretation: GDP began higher than GTP, but due to allosteric binding, GTP overtakes GDP.
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Hlavacek original model with Stites paper rates (gdat)
0.00E+00 5.00E+02 1.00E+03 1.50E+03 2.00E+03 2.50E+03 3.00E+03 3.50E+030.00E+00
5.00E+04
1.00E+05
1.50E+05
2.00E+05
2.50E+05
RasGTPRasGDP
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Hlavacek allosteric graph with converted rates (gdat)
0.00E+00 5.00E+02 1.00E+03 1.50E+03 2.00E+03 2.50E+03 3.00E+03 3.50E+030.00E+00
5.00E+04
1.00E+05
1.50E+05
2.00E+05
2.50E+05
RasGTPRasGDP
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Hlavacek model with converted rates, looking at increased Sos concentration
(scan)
0.00E+00 5.00E+09 1.00E+10 1.50E+10 2.00E+10 2.50E+100.00E+00
5.00E+04
1.00E+05
1.50E+05
2.00E+05
2.50E+05
3.00E+05
RasGTPRasGDP
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Minimal Model: units conversions & mutation effects
Replaced minimal model concentration values with Stites paper conversions to molecules
Mutations Removing GAP-mediated GTP hydrolysis produces a
simulation gdatLooked mainly at the G12V GAP insensitivity
mutation (Stites 2007)kf kr kcat
Other semi-tested mutations:Eliminated GAP (GAP-insensitive mutation)Removing the effector binding of Ras-GTP
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Initial: Minimal Model with GAP insensitivity mutation rates (gdat)
0.00E+00 5.00E+02 1.00E+03 1.50E+03 2.00E+03 2.50E+03 3.00E+03 3.50E+030.00E+00
1.00E+02
2.00E+02
3.00E+02
4.00E+02
5.00E+02
6.00E+02
7.00E+02
8.00E+02
9.00E+02
RasActRasGDP
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Further(1): Minimal Model with GAP insensitivity mutation rates (gdat)
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Further(2): Minimal Model with GAP insensitivity mutation rates (gdat)
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AcknowledgementsThank you to
Dr. Nancy Griffeth & NaralysDr. Jim Faeder NSF-CMACS Our fellow workshop members for collaboration!
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The End! Any Questions?
I’m about to run the
model guys!!!
Yea sure whatever
…
There he goes
again…