Amide Proton Exchange
-
Upload
clark-bartlett -
Category
Documents
-
view
52 -
download
2
description
Transcript of Amide Proton Exchange
![Page 1: Amide Proton Exchange](https://reader035.fdocuments.net/reader035/viewer/2022062321/56812cb2550346895d91665e/html5/thumbnails/1.jpg)
Amide Proton Exchange
![Page 2: Amide Proton Exchange](https://reader035.fdocuments.net/reader035/viewer/2022062321/56812cb2550346895d91665e/html5/thumbnails/2.jpg)
![Page 3: Amide Proton Exchange](https://reader035.fdocuments.net/reader035/viewer/2022062321/56812cb2550346895d91665e/html5/thumbnails/3.jpg)
![Page 4: Amide Proton Exchange](https://reader035.fdocuments.net/reader035/viewer/2022062321/56812cb2550346895d91665e/html5/thumbnails/4.jpg)
![Page 5: Amide Proton Exchange](https://reader035.fdocuments.net/reader035/viewer/2022062321/56812cb2550346895d91665e/html5/thumbnails/5.jpg)
![Page 6: Amide Proton Exchange](https://reader035.fdocuments.net/reader035/viewer/2022062321/56812cb2550346895d91665e/html5/thumbnails/6.jpg)
![Page 7: Amide Proton Exchange](https://reader035.fdocuments.net/reader035/viewer/2022062321/56812cb2550346895d91665e/html5/thumbnails/7.jpg)
![Page 8: Amide Proton Exchange](https://reader035.fdocuments.net/reader035/viewer/2022062321/56812cb2550346895d91665e/html5/thumbnails/8.jpg)
Amide Proton ExchangeProtection FactorF = kex,bound / kex,free
![Page 9: Amide Proton Exchange](https://reader035.fdocuments.net/reader035/viewer/2022062321/56812cb2550346895d91665e/html5/thumbnails/9.jpg)
![Page 10: Amide Proton Exchange](https://reader035.fdocuments.net/reader035/viewer/2022062321/56812cb2550346895d91665e/html5/thumbnails/10.jpg)
![Page 11: Amide Proton Exchange](https://reader035.fdocuments.net/reader035/viewer/2022062321/56812cb2550346895d91665e/html5/thumbnails/11.jpg)
![Page 12: Amide Proton Exchange](https://reader035.fdocuments.net/reader035/viewer/2022062321/56812cb2550346895d91665e/html5/thumbnails/12.jpg)
![Page 13: Amide Proton Exchange](https://reader035.fdocuments.net/reader035/viewer/2022062321/56812cb2550346895d91665e/html5/thumbnails/13.jpg)
![Page 14: Amide Proton Exchange](https://reader035.fdocuments.net/reader035/viewer/2022062321/56812cb2550346895d91665e/html5/thumbnails/14.jpg)
![Page 15: Amide Proton Exchange](https://reader035.fdocuments.net/reader035/viewer/2022062321/56812cb2550346895d91665e/html5/thumbnails/15.jpg)
SUPREXGhaemmaghami et al. PNAS 97: 8296 (2000)
![Page 16: Amide Proton Exchange](https://reader035.fdocuments.net/reader035/viewer/2022062321/56812cb2550346895d91665e/html5/thumbnails/16.jpg)
![Page 17: Amide Proton Exchange](https://reader035.fdocuments.net/reader035/viewer/2022062321/56812cb2550346895d91665e/html5/thumbnails/17.jpg)
![Page 18: Amide Proton Exchange](https://reader035.fdocuments.net/reader035/viewer/2022062321/56812cb2550346895d91665e/html5/thumbnails/18.jpg)
![Page 19: Amide Proton Exchange](https://reader035.fdocuments.net/reader035/viewer/2022062321/56812cb2550346895d91665e/html5/thumbnails/19.jpg)
Table 1. Stability of proteins by SUPREX and CD denaturationProtein Gf, kcal/mol Gf, kcal/mol SUPREXa CD SUPREXa CD 6-85 5.0 ± 0.4 4.4 ± 0.2b 6-85 (A66G) >4.0 >1.0 1.5c 6-85 (A63G) 4.8 ± 0.4 0.2 0.4c 6-85 (G46A/G48A/A66G) 5.2 ± 0.4 0.2 0.4c 6-85 (G46A/G48A/A49G) 5.6 ± 0.4 0.6 1.0c 6-85 (Q33Y) 6.5 ± 0.4 1.5 1.5d 6-85 (G46A/G48A) 6.7 ± 0.4 6.1 ± 0.2b 1.7 1.7 6-85 (G46A/G48A/Q33Y) 7.9 ± 0.7 2.9 3.4dMBP 16 ± 3 14.5 ± 0.4e
12.5 ± 0.2fMBP + 100 µM maltose 19 ± 3 3.0 2.9ga Stabilities measured at 23°C.b Stabilities measured at 25°C.c Gf (Gfvariant-Gfwild type) were measured at 37°C (23).d Gf measured at 25°C by G. Kapp, personal communication.e Stability measured at 25°C by urea titration (17).f Stability measured at 25°C by GdmCl titration (17).g A dissociation constant of 1 µM (20) and an estimated protein concentration of 5 µM were used to calculate the expected change in stability (3).