Talk:20.109(S13):Assess protein function (Day8)

Raw Data

Raw calcium titration data is posted below.

Day 7 data file, T/R	Day 7 data file, W/F
Media:S13-M2D7-TR-Red.txt	Media:S13-M2D7-WF-Red.txt
Media:S13-M2D7-TR-Org.txt	Media:S13-M2D7-WF-Org.txt
Media:S13-M2D7-TR-Ylw.txt	Media:S13-M2D7-WF-Ylw.txt
Media:S13-M2D7-TR-Grn.txt	Media:S13-M2D7-WF-Grn.txt
Media:S13-M2D7-TR-Blu.txt	Media:S13-M2D7-WF-Blu.txt
N/A	Media:S13-M2D7-WF-Pnk.txt
Media:S13-M2D7-TR-Prp.txt	Media:S13-M2D7-WF-Prp.txt
Media:S13-M2D7-TR-Plt.txt	Media:S13-M2D7-WF-Plt.txt

Analysis, T/R

Please be mindful of significant figures when you fill in this table. Do not state far more precise numbers than you can believe.

Group Colour	D24H, E67K, T79P, or M124S?	X#Z Mutant	Most believable KD; n for WT	Most believable KD; n for reference	Most believable KD; n for X#Z mutant	Comments (e.g., very low fluorescence)
Plt	M124S	D80R	WT: 0.0976; 1.4751	M124S: 0.4115; 3.9341	D80R: 1.2991; 2.8465	small difference between 0% and 100% for our mutant, affinity decrease and cooperativity increase
Orange	D24H	G25Q	0.1462, n = 0.8310	0.6759, n = 2.5734	0.7118; n = -0.2595	Very low fluorescence; oscillating trend...
Purple	M124S	M144R	KD = 0.0672, n = 1.1286	KD = 0.2682, n= 3.3655	KD = 0.3583, n= 4.1618	Higher cooperativity, lower affinity in target mutant
Blue	M124S	M71R	KD = 0.1175, n = 1.4880	KD = 0.3377, n = 3.3863	KD = 0.2246, n = 0.6741	Negative cooperativity, lower affinity in target mutant
Yellow	T79P	K77E	Kd = 0.1531, n = 1.4785	Kd = 0.8493, n = 1.5848	Kd = 0.0382, n = 1.1695	Design mutant: higher affinity, lower positive cooperativity
Green	T79P	M71R	Kd = 0.117, n = 1.50	Kd = 0.889, n = 4.05	Kd = 0.229, n = 0.684	T79P showed initial, slight increase in fluorescence, then sharp decrease. M71R showed sharply decreased cooperativity and increased Kd compared to WT.
Red	D24H	G25Q	Kd = 0.1232, n = 1.3583	Kd = 0.7541, n = 3.9963	Kd = 0.0415, n = -0.4091	Fluorescence rises to a peak then falls off at end

Analysis, W/F

Please be mindful of significant figures when you fill in this table. Do not state far more precise numbers than you can believe.

Group Colour	D24H, E67K, T79P, or M124S?	X#Z Mutant	Most believable KD; n for WT	Most believable KD; n for reference	Most believable KD; n for X#Z mutant	Comments (e.g., very low fluorescence)
Blue	D24H	D22E	Kd= 0.0781, n= 1.2420	Kd= 0.5804, n= 2.3194	Kd= 0.6807, n= 7.0509	Affinity decreased while the hill coefficient increased significantly. No extreme outliers.
Platinum	E67K	R126P	Kd = 0.0970 n =1.0639	Kd = 0.3729 n = 0.3830	Kd = 0.3488 n= 0.9085 Low fluorescent values
Pink	D24H	G25Q	Kd = 0.1232, n = 1.3583	Kd = 0.7541, n = 3.9963	Kd = 0.0415, n = -0.4091	Low fluorescence relative to wt, Affinity increased significantly or IPC was unable to refold properly.
Purple	M12S	M51T	Kd=0.1276, n=1.4726	Kd=0.3430, n=3.009	Kd=0.1191, n=1.2544
Yellow	M124S	M51T	Kd = 0.0739; n = 1.2038	Kd = 0.3859; n = 3.0814	Kd = 0.1403; n = 1.6126	Higher fluorescence compared to WT. Similar affinity as WT, higher affinity than reference.
Red	D24H	D22E	Kd = 0.0919, n = 1.858	Kd = 0.7099, n = 6.5600	Kd = 0.6134, n = 2.3619
Orange	D24H	D22E	Kd = 0.0916, n = 1.2896	Kd = 0.7357, n = 4.3991	Kd = 0.6766, n = 7.2874	Decreased affinity, increased cooperativity
Green	T79P	K77E	Kd = 6.75e-8M, n = 1.32	Kd = 9.30e-7M, n = 5.34	Kd = 1.84e-8M, n = 1.08