Talk:20.109(S12):Assess protein function (Day7)
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Raw Data
Please post your raw calcium titration data below.
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 0.1628; n 0.8206 for WT | KD 0.0520; n 0.2212 for E67K | KD 0.000047 for D20K | we used KDs from model 1 |
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. |