Talk:20.109(S13):Assess protein function (Day8): Difference between revisions
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|Design mutant: higher affinity, lower positive cooperativity | |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 | |||
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| Line 185: | Line 190: | ||
|K77E | |K77E | ||
|Kd = 6.75e-8M, n = 1.32 | |Kd = 6.75e-8M, n = 1.32 | ||
|Kd = 9.30e-7M, n = 5. | |Kd = 9.30e-7M, n = 5.34 | ||
|Kd = 1. | |Kd = 1.84e-8M, n = 1.08 | ||
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Latest revision as of 11:42, 24 April 2013
Raw Data
Raw calcium titration data is posted 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 | 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
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 |
