Talk:20.109(S12):Assess protein function (Day7)

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Raw Data

Please post your raw calcium titration data below.


Day 7 data file, T/R Day 7 data file, W/F
Media:S12_M2D7_TR-Red.txt Media:S12_M2D7_WF-Red.txt
Media:S12_M2D7_TR-Orange.txt Media:S12_M2D7_WF-Orange.txt
Media:S12_M2D7_TR-Yellow.txt Media:S12_M2D7_WF-Yellow.txt
Media:S12_M2D7_TR-Green.txt Media:S12_M2D7_WF-Green.txt
Media:S12_M2D7_TR-Blue.txt Media:S12_M2D7_WF-Blue.txt
Media:S12_M2D7_TR-Pink.txt Media:S12_M2D7_WF-Pink.txt
Media:S12_M2D7_TR-Purple.txt Media:S12_M2D7_WF-Purple.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 E67K, T79P, or M124S? X#Z Mutant Most believable KD; n for WT Most believable KD; n for (+) Most believable KD; n for X#Z mutant Comments
Orange T79P D24H 0.104, 1.26 0.166, 1.30 0.7892, 2.73 I utilized model 2.
Purple T79P F65D 0.0848, 1.313 0.1329, 1.3673 0.1923, 0.6862 Utilized model 2
Blue T79P D57N 0.0993, 1.1927 .1036, 1.4481 .0613, 1.2909 Utilized model 2
Yellow E67K D20R 0.0907, 1.2464 0.0300, 0.8097 0.6699, 0.4722 Utilize model 2
Green E67K E67R 0.0635, 1.945 0.0202, 0.9558 0.0279, 1.1453 Utilized model 2
Red E67K E67R 0.1429, 1.3167 0.0384, 0.9833 0.0600, 1.0852 Model 2
Pink T79P F65D 0.0877, 1.2104 0.1333, 1.3360 0.4273, 1.2646 Model 2

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 E67K, T79P, or M124S? X#Z Mutant Most believable KD; n for WT Most believable KD; n for (+) Most believable KD; n for X#Z mutant Comments
Orange T79P V55F KD 0.1025; n 1.8227 for WT KD 0.1491; n 1.3982 for T79P KD 0.1598; n 1.6114 for V55F The WT and T79P were switched during the experiment. For the above results, the labels were switched back to the original
Red T79P T79F KD 0.1075; n 1.3550 for WT KD 0.1372; n 1.1433 for T79P KD 0.1053; n 1.3550 for T79F As the above group, the labels were switched back to the original between T79P and WT. Also, our yield for WT and T79P was low (may be error prone)
Pink E67K D20K KD 0.0986; n 1.2648 for WT KD 0.0202; n 1.0027 for E67K KD 0.7043; n 0.3610 for D20K We used model 2 for our analysis. **Our data was updated in our revision process**
Purple M124S G23W/G25W KD 0.1643; n 1.3591 for WT KD 0.4753; n 2.9004 for M124S KD 0.1470; n 1.0503 for G23W/G25W Our yields for WT and G23W/G25W were low. The fluorescence values for G23W/G25W were very low (only between 18 and 24) and may not be significant. For G23W/G25W (contrary to WT and M124S), as [Ca2+] increased, fluorescence increased.
Blue E67K T26Y KD 0.1047; n 1.2123 KD 0.0193; n 0.8686 KD 0.3547; n -0.6402 We used KD and hill coefficient values from model 2. Our T26Y had minimal variance between our highest (58 A) and lowest (47 A) absorbance values, contributing to some noise -- was also had a reversed transition curve.
Green T79P F65D Method 1: KD 0.1151, Method 2:0.0872;Method 1: n 1.2658, Method 2: n = 1.1791 Method 1: KD 0.0969, Method 2:0.1018;Method 1: n 1.4356, Method 2: n = 1.6077 Method 1: KD 0.0599, Method 2:0.1523;Method 1: n 1.2302, Method 2: n = 1.4928 Results for both methods are as given.
Yellow E67K D20K KD 0.2155; n 1.6616 for WT KD 0.0556; n 1.0883 for E67K KD 0.0910; n 0.2797 for D20K We used the values from model 2. The Kd value and Hill coefficient for D20K should be ignored because from the raw data we can see that the fluorescence was basically flat, but

high enough to know it's not background.

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