[] = (deg cm2/dmol)

Protein + Protein + Protein + Protein +

Wavelength Native protein Denaturant A Denaturant B Denaturant C Denaturant D
[]

(nm) [] [] [] []

250 -6593.330523 -6745.901637 -6402.372556 -6765.355694 -358.1084677

249 -6755.88783 -6647.277859 -6610.685484 -6812.472874 -542.0032258
248 -6800.77175 -6494.052542 -6631.748534 -6825.012463 -435.3608871
247 -6936.022605 -6550.245601 -6664.835411 -6905.199413 -275.4903226
246 -7278.724096 -6718.394917 -6918.625122 -7064.300831 -172.5508065
245 -7662.85826 -6773.746212 -7304.898827 -7194.870357 -119.2173387
244 -8214.28739 -6794.949658 -7740.085777 -7338.701124 -262.108871
243 -8862.414101 -6933.858138 -8133.685606 -7671.704668 -384.8697581
242 -9388.270772 -7143.661901 -8481.992669 -8160.210288 -367.6245968
241 -10242.48473 -7352.860093 -9139.922898 -8634.503543 -505.0612903
240 -11561.34763 -7594.367424 -10091.24572 -9221.154814 -648.9717742
239 -12876.99902 -7875.610337 -10976.27126 -10034.27603 -615.4943548
238 -14457.21395 -8102.828568 -12126.13355 -10825.93194 -632.6294355
237 -16585.09592 -8285.943304 -13805.75941 -11625.06305 -676.1846774
236 -18962.27297 -8531.673754 -15615.15934 -12732.36461 -703.6891129
235 -21728.81232 -8866.388563 -17480.75415 -14070.2324 -858.5850806
234 -24793.0176 -9286.76674 -19536.60557 -15453.31122 -1010.339919
233 -27803.41606 -9716.245723 -21794.80462 -16936.08003 -1102.806855
232 -31160.14198 -10077.63783 -24393.59922 -18642.50929 -1352.252419
231 -34909.09519 -10397.38661 -27087.27016 -20477.99756 -1589.717742
230 -38654.67326 -10659.16459 -29610.59409 -22113.29069 -1531.415726
229 -42463.85899 -10905.06085 -32297.9306 -23574.90347 -1509.696371
228 -46068.84176 -11218.64614 -35004.08834 -25225.76686 -1617.045565
227 -49173.07564 -11575.18732 -37228.14064 -27012.71909 -1482.729032
226 -52284.31635 -11942.71078 -39323.11058 -28506.05609 -1551.450403
225 -55442.04093 -12192.27884 -41515.451 -29529.53275 -2055.82621
224 -57696.08346 -12279.77627 -43287.09005 -30390.136 -2033.504839
223 -58951.37463 -12409.59922 -44513.10606 -31283.47581 -1530.909677
222 -60297.86364 -12493.7781 -45492.19795 -31918.30718 -1663.037097
221 -61986.43671 -12306.85533 -46446.78715 -32270.91129 -2161.84879
220 -63223.30853 -12125.45711 -47308.76527 -32586.60716 -2015.431048
219 -63660.03519 -12263.12854 -47642.79374 -32806.59433 -1652.960484
218 -63685.52346 -12468.80682 -47553.48558 -32960.40286 -2062.633065
217 -63691.55144 -12478.23473 -47673.09054 -33076.26527 -2590.0125
216 -63952.13551 -12767.12378 -48023.7665 -33192.33077 -2304.721774
215 -64939.71554 -13639.80511 -48466.42229 -33721.91239 -2007.22621
214 -66297.40958 -14132.42363 -49249.22923 -34563.86376 -2272.445565
213 -67392.07649 -13919.33419 -50258.76466 -35403.40848 -2828.600806
212 -69243.69 -14327.95797 -51588.09836 -36657.06281 -3685.87379
211 -72726.96481 -15657.19135 -54144.78372 -38526.76686 -3914.544758
210 -77408.7065 -17226.34861 -57857.86657 -40838.0793 -3261.722581
209 -82548.96334 -19135.34739 -61691.31549 -43437.43121 -3725.81371
208 -86925.85288 -20971.201 -65122.51613 -45999.75379 -5400.914113
207 -89865.84836 -22488.93597 -67201.53482 -48107.10044 -6459.911694
206 -90670.8238 -24668.23766 -67197.80205 -48936.16435 -7280.825403
205 -87536.76576 -27396.38038 -65273.97959 -48185.85472 -8742.503629
204 -80419.38135 -29832.10288 -60906.38258 -46218.95699 -10400.75524
203 -71195.47397 -31898.47361 -53941.48082 -42482.92864 -12236.10605
202 -59572.88832 -33374.01478 -44454.32307 -37040.16923 -13720.30242
201 -43100.69966 -34098.8161 -32242.61217 -30650.84348 -13919.31411
200 -22391.17901 -34459.78825 -18236.33859 -22702.95051 -14459.97419
199 89.98680352 -35026.55425 -1970.48827 -13092.20674 -16653.13992
198 24952.64919 -35388.06574 17163.48277 -2215.214565 -18000.07298
197 51805.80584 -33849.40909 37540.4053 10673.37781 -17298.23548
196 78576.63649 -30571.06317 58117.20125 24717.06855 -16545.93831
195 101174.5479 -27596.03702 74787.10459 35724.72715 -15903.38629
194 111047.4395 -24726.53922 83544.10667 40434.49523 -14485.33589
193 111283.0051 -19899.3563 88564.42278 42446.86339 -12893.95927
192 111462.8834 -14118.39137 92128.35875 46855.29191 -11873.9379
191 103204.4961 -13080.66178 93202.74976 50516.52102 -12820.89153
190 85162.88661 -18840.22752 95842.13563 50158.13368 -16132.2004
 CD Spectroscopy

plot molar ellip vs lambda

molar ellipticity vs wa

native protein protein + denaturant A protein + denaturant B

can you say that your protein is optically active?

CD can only measure optically active (with stereogenic C, chirality

Viscometry: TNR 11, attach table label which is best denaturant

CD Spectro
plot theta vs lambda
explain what 2" structure is dominant
(effect of denaturant - intensity of absorbance decreases)
 negative absorbance: direction of rotation (arbitrary but it matters

determine delta epsilon*

llipticity vs wavelength

protein + denaturant B protein + denaturant C protein + denaturant D

does this have alpha helices? beta sheets?

reogenic C, chirality) (what values)
(albumin; prove that it is alpha helix)

denaturant
bitrary but it matters lol)
 Protein + Protein + Protein + Protein + Protein +
Wavelength
Denaturant A Denaturant B Denaturant C Denaturant D Denaturant A

(nm) [] [] [] [] ()

250 -6745.901637 -6402.372556 -6765.355694 -358.1084677 -2.04532825

249 -6647.277859 -6610.685484 -6812.472874 -542.0032258 -2.01542595
248 -6494.052542 -6631.748534 -6825.012463 -435.3608871 -1.96896869
247 -6550.245601 -6664.835411 -6905.199413 -275.4903226 -1.98600619
246 -6718.394917 -6918.625122 -7064.300831 -172.5508065 -2.03698833
245 -6773.746212 -7304.898827 -7194.870357 -119.2173387 -2.05377061
244 -6794.949658 -7740.085777 -7338.701124 -262.108871 -2.0601994
243 -6933.858138 -8133.685606 -7671.704668 -384.8697581 -2.10231585
242 -7143.661901 -8481.992669 -8160.210288 -367.6245968 -2.16592745
241 -7352.860093 -9139.922898 -8634.503543 -505.0612903 -2.22935543
240 -7594.367424 -10091.24572 -9221.154814 -648.9717742 -2.30257941
239 -7875.610337 -10976.27126 -10034.27603 -615.4943548 -2.38785105
238 -8102.828568 -12126.13355 -10825.93194 -632.6294355 -2.45674264
237 -8285.943304 -13805.75941 -11625.06305 -676.1846774 -2.51226224
236 -8531.673754 -15615.15934 -12732.36461 -703.6891129 -2.58676665
235 -8866.388563 -17480.75415 -14070.2324 -858.5850806 -2.68825073
234 -9286.76674 -19536.60557 -15453.31122 -1010.339919 -2.81570758
233 -9716.245723 -21794.80462 -16936.08003 -1102.806855 -2.94592375
232 -10077.63783 -24393.59922 -18642.50929 -1352.252419 -3.05549628
231 -10397.38661 -27087.27016 -20477.99756 -1589.717742 -3.15244273
230 -10659.16459 -29610.59409 -22113.29069 -1531.415726 -3.23181268
229 -10905.06085 -32297.9306 -23574.90347 -1509.696371 -3.30636737
228 -11218.64614 -35004.08834 -25225.76686 -1617.045565 -3.40144507
227 -11575.18732 -37228.14064 -27012.71909 -1482.729032 -3.50954682
226 -11942.71078 -39323.11058 -28506.05609 -1551.450403 -3.62097835
225 -12192.27884 -41515.451 -29529.53275 -2055.82621 -3.6966463
224 -12279.77627 -43287.09005 -30390.136 -2033.504839 -3.72317515
223 -12409.59922 -44513.10606 -31283.47581 -1530.909677 -3.7625369
222 -12493.7781 -45492.19795 -31918.30718 -1663.037097 -3.78805958
221 -12306.85533 -46446.78715 -32270.91129 -2161.84879 -3.7313854
220 -12125.45711 -47308.76527 -32586.60716 -2015.431048 -3.67638624
219 -12263.12854 -47642.79374 -32806.59433 -1652.960484 -3.71812763
218 -12468.80682 -47553.48558 -32960.40286 -2062.633065 -3.78048839
217 -12478.23473 -47673.09054 -33076.26527 -2590.0125 -3.7833469
216 -12767.12378 -48023.7665 -33192.33077 -2304.721774 -3.87093681
215 -13639.80511 -48466.42229 -33721.91239 -2007.22621 -4.13553002
214 -14132.42363 -49249.22923 -34563.86376 -2272.445565 -4.28488983
213 -13919.33419 -50258.76466 -35403.40848 -2828.600806 -4.22028203
212 -14327.95797 -51588.09836 -36657.06281 -3685.87379 -4.344175
211 -15657.19135 -54144.78372 -38526.76686 -3914.544758 -4.74719282
210 -17226.34861 -57857.86657 -40838.0793 -3261.722581 -5.22295452
209 -19135.34739 -61691.31549 -43437.43121 -3725.81371 -5.80175471
208 -20971.201 -65122.51613 -45999.75379 -5400.914113 -6.3583776
207 -22488.93597 -67201.53482 -48107.10044 -6459.911694 -6.81854829
206 -24668.23766 -67197.80205 -48936.16435 -7280.825403 -7.47930315
205 -27396.38038 -65273.97959 -48185.85472 -8742.503629 -8.30646425
204 -29832.10288 -60906.38258 -46218.95699 -10400.75524 -9.04496479
203 -31898.47361 -53941.48082 -42482.92864 -12236.10605 -9.67147948
202 -33374.01478 -44454.32307 -37040.16923 -13720.30242 -10.1188572
201 -34098.8161 -32242.61217 -30650.84348 -13919.31411 -10.3386138
200 -34459.78825 -18236.33859 -22702.95051 -14459.97419 -10.448059
199 -35026.55425 -1970.48827 -13092.20674 -16653.13992 -10.6199
198 -35388.06574 17163.48277 -2215.214565 -18000.07298 -10.7295087
197 -33849.40909 37540.4053 10673.37781 -17298.23548 -10.2629947
196 -30571.06317 58117.20125 24717.06855 -16545.93831 -9.26901436
195 -27596.03702 74787.10459 35724.72715 -15903.38629 -8.36699928
194 -24726.53922 83544.10667 40434.49523 -14485.33589 -7.49697993
193 -19899.3563 88564.42278 42446.86339 -12893.95927 -6.03339891
192 -14118.39137 92128.35875 46855.29191 -11873.9379 -4.28063531
191 -13080.66178 93202.74976 50516.52102 -12820.89153 -3.96600018
190 -18840.22752 95842.13563 50158.13368 -16132.2004 -5.71227564

C/O http://perry.freeshell.org/raussens.html
A B
Helix(%):5.2 Helix(%):35.8
 Beta(%):36.0 Beta(%):13.6
Turn(%):12.5 Turn(%):12.5
Random(%):41.2 Random(%):33.8

Sum(%):95.0 Sum(%):95.7

A B C
250 -2.04532825 250 -1.94117172 250
249 -2.01542595 249 -2.0043313 249
248 -1.96896869 248 -2.01071752 248
247 -1.98600619 247 -2.02074932 247
246 -2.03698833 246 -2.09769727 246
245 -2.05377061 245 -2.21481379 245
244 -2.0601994 244 -2.34676059 244
243 -2.10231585 243 -2.46609836 243
242 -2.16592745 242 -2.57170356 242
241 -2.22935543 241 -2.77118516 241
240 -2.30257941 240 -3.05962213 240
239 -2.38785105 239 -3.32795806 239
238 -2.45674264 238 -3.67659134 238
237 -2.51226224 237 -4.18584665 237
236 -2.58676665 236 -4.73444889 236
235 -2.68825073 235 -5.30008919 235
234 -2.81570758 234 -5.92341446 234
233 -2.94592375 233 -6.60809066 233
232 -3.05549628 232 -7.39603396 232
231 -3.15244273 231 -8.21274336 231
230 -3.23181268 230 -8.97780428 230
229 -3.30636737 229 -9.79259311 229
228 -3.40144507 228 -10.6130885 228
227 -3.50954682 227 -11.2874115 227
226 -3.62097835 226 -11.9225974 226
225 -3.6966463 225 -12.5873055 225
224 -3.72317515 224 -13.1244588 224
223 -3.7625369 223 -13.4961816 223
222 -3.78805958 222 -13.793038 222
221 -3.7313854 221 -14.0824653 221
220 -3.67638624 220 -14.3438134 220
219 -3.71812763 219 -14.4450894 219
218 -3.78048839 218 -14.4180115 218
217 -3.7833469 217 -14.4542752 217
216 -3.87093681 216 -14.5605987 216
215 -4.13553002 215 -14.69481 215
214 -4.28488983 214 -14.9321537 214
213 -4.22028203 213 -15.2382405 213
212 -4.344175 212 -15.6412887 212
211 -4.74719282 211 -16.4164647 211
210 -5.22295452 210 -17.5422553 210
209 -5.80175471 209 -18.7045405 209
208 -6.3583776 208 -19.7448657 208
207 -6.81854829 207 -20.3752152 207
206 -7.47930315 206 -20.3740835 206
205 -8.30646425 205 -19.7907888 205
204 -9.04496479 204 -18.4665522 204
203 -9.67147948 203 -16.3548241 203
202 -10.1188572 202 -13.4783588 202
201 -10.3386138 201 -9.7758208 201
200 -10.448059 200 -5.52917912 200
199 -10.6199 199 -0.59744354 199
198 -10.7295087 198 5.20389387 198
197 -10.2629947 197 11.3820888 197
196 -9.26901436 196 17.6208845 196
195 -8.36699928 195 22.6751272 195
194 -7.49697993 194 25.3302124 194
193 -6.03339891 193 26.8523506 193
192 -4.28063531 192 27.9329206 192
191 -3.96600018 191 28.2586713 191
190 -5.71227564 190 29.0589217 190
 Protein + Protein + Protein +
Denaturant B Denaturant C Denaturant D

() () ()

-1.94117172 -2.05122664 -0.10857694 DELTA EPSILON IS UNIT OF ABSORBANCE

-2.0043313 -2.06551236 -0.16433304 WHATS THE POINT
-2.01071752 -2.06931431 -0.13199954
-2.02074932 -2.09362665 -0.08352748 det 2" struc
-2.09769727 -2.14186551 -0.05231666 know best denaturant
-2.21481379 -2.18145363 -0.03614618 get delta E
-2.34676059 -2.2250625 -0.07947028 use it to get %2" structure
-2.46609836 -2.32602773 -0.11669085 search K2D3. click the first thing
-2.57170356 -2.47414053 -0.11146219 wavelength: 190 to 250 nm
-2.77118516 -2.61794419 -0.1531324 CD Units delta epsilon
-3.05962213 -2.79581433 -0.19676544 protein size leave blank
-3.32795806 -3.04234917 -0.18661523 CD spectrum: delta epsilon
-3.67659134 -3.28237582 -0.19181051 * higher percent 2" less den
-4.18584665 -3.52466893 -0.20501627 trying to monitor conformation thru visc
-4.73444889 -3.86039798 -0.2133555
-5.30008919 -4.26603372 -0.26031929 * may include figure in discussion, but p
-5.92341446 -4.68537724 -0.3063307
-6.60809066 -5.13494634 -0.33436628
-7.39603396 -5.65232833 -0.4099971
-8.21274336 -6.20884045 -0.48199556
-8.97780428 -6.70465426 -0.46431864
-9.79259311 -7.14780895 -0.45773342
-10.6130885 -7.6483436 -0.49028123
-11.2874115 -8.1901398 -0.44955704
-11.9225974 -8.64291313 -0.47039306
-12.5873055 -8.95322684 -0.62331763
-13.1244588 -9.21415803 -0.61654989
-13.4961816 -9.4850148 -0.4641652
-13.793038 -9.67749293 -0.50422567
-14.0824653 -9.78440097 -0.65546322
-14.3438134 -9.8801186 -0.61106999
-14.4450894 -9.94681776 -0.50117048
-14.4180115 -9.99345184 -0.62538144
-14.4542752 -10.0285808 -0.78528061
-14.5605987 -10.0637714 -0.69878169
-14.69481 -10.2243382 -0.60858232
-14.9321537 -10.4796143 -0.68899568
-15.2382405 -10.7341606 -0.85761955
-15.6412887 -11.1142632 -1.11754102
-16.4164647 -11.6811494 -1.18687307
-17.5422553 -12.3819293 -0.9889402
-18.7045405 -13.1700416 -1.12965063
-19.7448657 -13.9469267 -1.63753384
-20.3752152 -14.5858652 -1.95861733
-20.3740835 -14.8372337 -2.20751483
-19.7907888 -14.6097431 -2.65068935
-18.4665522 -14.0133882 -3.15346408
-16.3548241 -12.8806405 -3.70993452
-13.4783588 -11.2304194 -4.15993646
-9.7758208 -9.29320341 -4.22027594
-5.52917912 -6.88343657 -4.38420174
-0.59744354 -3.96950056 -5.04916012
5.20389387 -0.67164349 -5.45754441
11.3820888 3.23612207 -5.24475031
17.6208845 7.49410847 -5.01665706
22.6751272 10.831583 -4.82183806
25.3302124 12.2595644 -4.3918913
26.8523506 12.8697057 -3.90939278
27.9329206 14.2063222 -3.60012671
28.2586713 15.3163911 -3.88723896
29.0589217 15.2077296 -4.89121351

C D
Helix(%):27.8 Helix(%):2.7
Beta(%):18.1 Beta(%):44.2
Turn(%):12.5 Turn(%):12.5
Random(%):35.7 Random(%):42.9

Sum(%):94.1 Sum(%):96.9

D
-2.05122664 250 -0.10857694
-2.06551236 249 -0.16433304
-2.06931431 248 -0.13199954
-2.09362665 247 -0.08352748
-2.14186551 246 -0.05231666
-2.18145363 245 -0.03614618
-2.2250625 244 -0.07947028
-2.32602773 243 -0.11669085
-2.47414053 242 -0.11146219
-2.61794419 241 -0.1531324
-2.79581433 240 -0.19676544
-3.04234917 239 -0.18661523
-3.28237582 238 -0.19181051
-3.52466893 237 -0.20501627
-3.86039798 236 -0.2133555
-4.26603372 235 -0.26031929
-4.68537724 234 -0.3063307
-5.13494634 233 -0.33436628
-5.65232833 232 -0.4099971
-6.20884045 231 -0.48199556
-6.70465426 230 -0.46431864
-7.14780895 229 -0.45773342
-7.6483436 228 -0.49028123
-8.1901398 227 -0.44955704
-8.64291313 226 -0.47039306
-8.95322684 225 -0.62331763
-9.21415803 224 -0.61654989
-9.4850148 223 -0.4641652
-9.67749293 222 -0.50422567
-9.78440097 221 -0.65546322
-9.8801186 220 -0.61106999
-9.94681776 219 -0.50117048
-9.99345184 218 -0.62538144
-10.0285808 217 -0.78528061
-10.0637714 216 -0.69878169
-10.2243382 215 -0.60858232
-10.4796143 214 -0.68899568
-10.7341606 213 -0.85761955
-11.1142632 212 -1.11754102
-11.6811494 211 -1.18687307
-12.3819293 210 -0.9889402
-13.1700416 209 -1.12965063
-13.9469267 208 -1.63753384
-14.5858652 207 -1.95861733
-14.8372337 206 -2.20751483
-14.6097431 205 -2.65068935
-14.0133882 204 -3.15346408
-12.8806405 203 -3.70993452
-11.2304194 202 -4.15993646
-9.29320341 201 -4.22027594
-6.88343657 200 -4.38420174
-3.96950056 199 -5.04916012
-0.67164349 198 -5.45754441
3.23612207 197 -5.24475031
7.49410847 196 -5.01665706
10.831583 195 -4.82183806
12.2595644 194 -4.3918913
12.8697057 193 -3.90939278
14.2063222 192 -3.60012671
15.3163911 191 -3.88723896
15.2077296 190 -4.89121351
 UNIT OF ABSORBANCE????????

k the first thing

elength: 190 to 250 nm
Units delta epsilon
ein size leave blank
pectrum: delta epsilon values
er percent 2" less denatured. best denaturant has low helicity
conformation thru viscosity and CD spectroscopy

re in discussion, but put in appendix

A B C
190 -5.71227564 190 29.0589217 190 15.2077296
191 -3.96600018 191 28.2586713 191 15.3163911
192 -4.28063531 192 27.9329206 192 14.2063222
193 -6.03339891 193 26.8523506 193 12.8697057
194 -7.49697993 194 25.3302124 194 12.2595644
195 -8.36699928 195 22.6751272 195 10.831583
196 -9.26901436 196 17.6208845 196 7.49410847
197 -10.2629947 197 11.3820888 197 3.23612207
198 -10.7295087 198 5.20389387 198 -0.67164349
199 -10.6199 199 -0.59744354 199 -3.96950056
200 -10.448059 200 -5.52917912 200 -6.88343657
201 -10.3386138 201 -9.7758208 201 -9.29320341
202 -10.1188572 202 -13.4783588 202 -11.2304194
203 -9.67147948 203 -16.3548241 203 -12.8806405
204 -9.04496479 204 -18.4665522 204 -14.0133882
205 -8.30646425 205 -19.7907888 205 -14.6097431
206 -7.47930315 206 -20.3740835 206 -14.8372337
207 -6.81854829 207 -20.3752152 207 -14.5858652
208 -6.3583776 208 -19.7448657 208 -13.9469267
209 -5.80175471 209 -18.7045405 209 -13.1700416
210 -5.22295452 210 -17.5422553 210 -12.3819293
211 -4.74719282 211 -16.4164647 211 -11.6811494
212 -4.344175 212 -15.6412887 212 -11.1142632
213 -4.22028203 213 -15.2382405 213 -10.7341606
214 -4.28488983 214 -14.9321537 214 -10.4796143
215 -4.13553002 215 -14.69481 215 -10.2243382
216 -3.87093681 216 -14.5605987 216 -10.0637714
217 -3.7833469 217 -14.4542752 217 -10.0285808
218 -3.78048839 218 -14.4180115 218 -9.99345184
219 -3.71812763 219 -14.4450894 219 -9.94681776
220 -3.67638624 220 -14.3438134 220 -9.8801186
221 -3.7313854 221 -14.0824653 221 -9.78440097
222 -3.78805958 222 -13.793038 222 -9.67749293
223 -3.7625369 223 -13.4961816 223 -9.4850148
224 -3.72317515 224 -13.1244588 224 -9.21415803
225 -3.6966463 225 -12.5873055 225 -8.95322684
226 -3.62097835 226 -11.9225974 226 -8.64291313
227 -3.50954682 227 -11.2874115 227 -8.1901398
228 -3.40144507 228 -10.6130885 228 -7.6483436
229 -3.30636737 229 -9.79259311 229 -7.14780895
230 -3.23181268 230 -8.97780428 230 -6.70465426
231 -3.15244273 231 -8.21274336 231 -6.20884045
232 -3.05549628 232 -7.39603396 232 -5.65232833
233 -2.94592375 233 -6.60809066 233 -5.13494634
234 -2.81570758 234 -5.92341446 234 -4.68537724
235 -2.68825073 235 -5.30008919 235 -4.26603372
236 -2.58676665 236 -4.73444889 236 -3.86039798
237 -2.51226224 237 -4.18584665 237 -3.52466893
238 -2.45674264 238 -3.67659134 238 -3.28237582
239 -2.38785105 239 -3.32795806 239 -3.04234917
240 -2.30257941 240 -3.05962213 240 -2.79581433
D
190 -4.89121351
191 -3.88723896
192 -3.60012671
193 -3.90939278
194 -4.3918913
195 -4.82183806
196 -5.01665706
197 -5.24475031
198 -5.45754441
199 -5.04916012
200 -4.38420174
201 -4.22027594
202 -4.15993646
203 -3.70993452
204 -3.15346408
205 -2.65068935
206 -2.20751483
207 -1.95861733
208 -1.63753384
209 -1.12965063
210 -0.9889402
211 -1.18687307
212 -1.11754102
213 -0.85761955
214 -0.68899568
215 -0.60858232
216 -0.69878169
217 -0.78528061
218 -0.62538144
219 -0.50117048
220 -0.61106999
221 -0.65546322
222 -0.50422567
223 -0.4641652
224 -0.61654989
225 -0.62331763
226 -0.47039306
227 -0.44955704
228 -0.49028123
229 -0.45773342
230 -0.46431864
231 -0.48199556
232 -0.4099971
233 -0.33436628
234 -0.3063307
235 -0.26031929
236 -0.2133555
237 -0.20501627
238 -0.19181051
239 -0.18661523
240 -0.19676544
 [] = (deg cm2/dmol)

Protein + Protein + Protein + Protein +

Wavelength Native protein Denaturant A Denaturant B Denaturant C Denaturant D
[]

(nm) [] [] [] []

250 -6593.330523 -6745.901637 -6402.372556 -6765.355694 -358.1084677

249 -6755.88783 -6647.277859 -6610.685484 -6812.472874 -542.0032258
248 -6800.77175 -6494.052542 -6631.748534 -6825.012463 -435.3608871
247 -6936.022605 -6550.245601 -6664.835411 -6905.199413 -275.4903226
246 -7278.724096 -6718.394917 -6918.625122 -7064.300831 -172.5508065
245 -7662.85826 -6773.746212 -7304.898827 -7194.870357 -119.2173387
244 -8214.28739 -6794.949658 -7740.085777 -7338.701124 -262.108871
243 -8862.414101 -6933.858138 -8133.685606 -7671.704668 -384.8697581
242 -9388.270772 -7143.661901 -8481.992669 -8160.210288 -367.6245968
241 -10242.48473 -7352.860093 -9139.922898 -8634.503543 -505.0612903
240 -11561.34763 -7594.367424 -10091.24572 -9221.154814 -648.9717742
239 -12876.99902 -7875.610337 -10976.27126 -10034.27603 -615.4943548
238 -14457.21395 -8102.828568 -12126.13355 -10825.93194 -632.6294355
237 -16585.09592 -8285.943304 -13805.75941 -11625.06305 -676.1846774
236 -18962.27297 -8531.673754 -15615.15934 -12732.36461 -703.6891129
235 -21728.81232 -8866.388563 -17480.75415 -14070.2324 -858.5850806
234 -24793.0176 -9286.76674 -19536.60557 -15453.31122 -1010.339919
233 -27803.41606 -9716.245723 -21794.80462 -16936.08003 -1102.806855
232 -31160.14198 -10077.63783 -24393.59922 -18642.50929 -1352.252419
231 -34909.09519 -10397.38661 -27087.27016 -20477.99756 -1589.717742
230 -38654.67326 -10659.16459 -29610.59409 -22113.29069 -1531.415726
229 -42463.85899 -10905.06085 -32297.9306 -23574.90347 -1509.696371
228 -46068.84176 -11218.64614 -35004.08834 -25225.76686 -1617.045565
227 -49173.07564 -11575.18732 -37228.14064 -27012.71909 -1482.729032
226 -52284.31635 -11942.71078 -39323.11058 -28506.05609 -1551.450403
225 -55442.04093 -12192.27884 -41515.451 -29529.53275 -2055.82621
224 -57696.08346 -12279.77627 -43287.09005 -30390.136 -2033.504839
223 -58951.37463 -12409.59922 -44513.10606 -31283.47581 -1530.909677
222 -60297.86364 -12493.7781 -45492.19795 -31918.30718 -1663.037097
221 -61986.43671 -12306.85533 -46446.78715 -32270.91129 -2161.84879
220 -63223.30853 -12125.45711 -47308.76527 -32586.60716 -2015.431048
219 -63660.03519 -12263.12854 -47642.79374 -32806.59433 -1652.960484
218 -63685.52346 -12468.80682 -47553.48558 -32960.40286 -2062.633065
217 -63691.55144 -12478.23473 -47673.09054 -33076.26527 -2590.0125
216 -63952.13551 -12767.12378 -48023.7665 -33192.33077 -2304.721774
215 -64939.71554 -13639.80511 -48466.42229 -33721.91239 -2007.22621
214 -66297.40958 -14132.42363 -49249.22923 -34563.86376 -2272.445565
213 -67392.07649 -13919.33419 -50258.76466 -35403.40848 -2828.600806
212 -69243.69 -14327.95797 -51588.09836 -36657.06281 -3685.87379
211 -72726.96481 -15657.19135 -54144.78372 -38526.76686 -3914.544758
210 -77408.7065 -17226.34861 -57857.86657 -40838.0793 -3261.722581
209 -82548.96334 -19135.34739 -61691.31549 -43437.43121 -3725.81371
208 -86925.85288 -20971.201 -65122.51613 -45999.75379 -5400.914113
207 -89865.84836 -22488.93597 -67201.53482 -48107.10044 -6459.911694
206 -90670.8238 -24668.23766 -67197.80205 -48936.16435 -7280.825403
205 -87536.76576 -27396.38038 -65273.97959 -48185.85472 -8742.503629
204 -80419.38135 -29832.10288 -60906.38258 -46218.95699 -10400.75524
203 -71195.47397 -31898.47361 -53941.48082 -42482.92864 -12236.10605
202 -59572.88832 -33374.01478 -44454.32307 -37040.16923 -13720.30242
201 -43100.69966 -34098.8161 -32242.61217 -30650.84348 -13919.31411
200 -22391.17901 -34459.78825 -18236.33859 -22702.95051 -14459.97419
199 89.98680352 -35026.55425 -1970.48827 -13092.20674 -16653.13992
198 24952.64919 -35388.06574 17163.48277 -2215.214565 -18000.07298
197 51805.80584 -33849.40909 37540.4053 10673.37781 -17298.23548
196 78576.63649 -30571.06317 58117.20125 24717.06855 -16545.93831
195 101174.5479 -27596.03702 74787.10459 35724.72715 -15903.38629
194 111047.4395 -24726.53922 83544.10667 40434.49523 -14485.33589
193 111283.0051 -19899.3563 88564.42278 42446.86339 -12893.95927
192 111462.8834 -14118.39137 92128.35875 46855.29191 -11873.9379
191 103204.4961 -13080.66178 93202.74976 50516.52102 -12820.89153
190 85162.88661 -18840.22752 95842.13563 50158.13368 -16132.2004
 CD Spectroscopy

plot molar ellip vs lambda

molar ellipticity vs wa
150000

100000

50000

molar ellipticity 0

-50000

-100000

-150000

native protein protein + denaturant A protein + denaturant B

can you say that your protein is optically active?

CD can only measure optically active (with stereogenic C, chirality

Viscometry: TNR 11, attach table label which is best denaturant

CD Spectro
plot theta vs lambda
explain what 2" structure is dominant
(effect of denaturant - intensity of absorbance decreases)
 negative absorbance: direction of rotation (arbitrary but it matters

determine delta epsilon*

llipticity vs wavelength

protein + denaturant B protein + denaturant C protein + denaturant D

does this have alpha helices? beta sheets?

reogenic C, chirality) (what values)
(albumin; prove that it is alpha helix)

denaturant
bitrary but it matters lol)
 delta e
Protein + Protein + Protein + Protein +
Wavelength A
Denaturant A Denaturant B Denaturant C Denaturant D

(nm) [] [] [] []

250 -6745.901637 -6402.372556 -6765.355694 -358.1084677 -2.04532825

249 -6647.277859 -6610.685484 -6812.472874 -542.0032258 -2.01542595
248 -6494.052542 -6631.748534 -6825.012463 -435.3608871 -1.96896869
247 -6550.245601 -6664.835411 -6905.199413 -275.4903226 -1.98600619
246 -6718.394917 -6918.625122 -7064.300831 -172.5508065 -2.03698833
245 -6773.746212 -7304.898827 -7194.870357 -119.2173387 -2.05377061
244 -6794.949658 -7740.085777 -7338.701124 -262.108871 -2.0601994
243 -6933.858138 -8133.685606 -7671.704668 -384.8697581 -2.10231585
242 -7143.661901 -8481.992669 -8160.210288 -367.6245968 -2.16592745
241 -7352.860093 -9139.922898 -8634.503543 -505.0612903 -2.22935543
240 -7594.367424 -10091.24572 -9221.154814 -648.9717742 -2.30257941
239 -7875.610337 -10976.27126 -10034.27603 -615.4943548 -2.38785105
238 -8102.828568 -12126.13355 -10825.93194 -632.6294355 -2.45674264
237 -8285.943304 -13805.75941 -11625.06305 -676.1846774 -2.51226224
236 -8531.673754 -15615.15934 -12732.36461 -703.6891129 -2.58676665
235 -8866.388563 -17480.75415 -14070.2324 -858.5850806 -2.68825073
234 -9286.76674 -19536.60557 -15453.31122 -1010.339919 -2.81570758
233 -9716.245723 -21794.80462 -16936.08003 -1102.806855 -2.94592375
232 -10077.63783 -24393.59922 -18642.50929 -1352.252419 -3.05549628
231 -10397.38661 -27087.27016 -20477.99756 -1589.717742 -3.15244273
230 -10659.16459 -29610.59409 -22113.29069 -1531.415726 -3.23181268
229 -10905.06085 -32297.9306 -23574.90347 -1509.696371 -3.30636737
228 -11218.64614 -35004.08834 -25225.76686 -1617.045565 -3.40144507
227 -11575.18732 -37228.14064 -27012.71909 -1482.729032 -3.50954682
226 -11942.71078 -39323.11058 -28506.05609 -1551.450403 -3.62097835
225 -12192.27884 -41515.451 -29529.53275 -2055.82621 -3.6966463
224 -12279.77627 -43287.09005 -30390.136 -2033.504839 -3.72317515
223 -12409.59922 -44513.10606 -31283.47581 -1530.909677 -3.7625369
222 -12493.7781 -45492.19795 -31918.30718 -1663.037097 -3.78805958
221 -12306.85533 -46446.78715 -32270.91129 -2161.84879 -3.7313854
220 -12125.45711 -47308.76527 -32586.60716 -2015.431048 -3.67638624
219 -12263.12854 -47642.79374 -32806.59433 -1652.960484 -3.71812763
218 -12468.80682 -47553.48558 -32960.40286 -2062.633065 -3.78048839
217 -12478.23473 -47673.09054 -33076.26527 -2590.0125 -3.7833469
216 -12767.12378 -48023.7665 -33192.33077 -2304.721774 -3.87093681
215 -13639.80511 -48466.42229 -33721.91239 -2007.22621 -4.13553002
214 -14132.42363 -49249.22923 -34563.86376 -2272.445565 -4.28488983
213 -13919.33419 -50258.76466 -35403.40848 -2828.600806 -4.22028203
212 -14327.95797 -51588.09836 -36657.06281 -3685.87379 -4.344175
211 -15657.19135 -54144.78372 -38526.76686 -3914.544758 -4.74719282
210 -17226.34861 -57857.86657 -40838.0793 -3261.722581 -5.22295452
209 -19135.34739 -61691.31549 -43437.43121 -3725.81371 -5.80175471
208 -20971.201 -65122.51613 -45999.75379 -5400.914113 -6.3583776
207 -22488.93597 -67201.53482 -48107.10044 -6459.911694 -6.81854829
206 -24668.23766 -67197.80205 -48936.16435 -7280.825403 -7.47930315
205 -27396.38038 -65273.97959 -48185.85472 -8742.503629 -8.30646425
204 -29832.10288 -60906.38258 -46218.95699 -10400.75524 -9.04496479
203 -31898.47361 -53941.48082 -42482.92864 -12236.10605 -9.67147948
202 -33374.01478 -44454.32307 -37040.16923 -13720.30242 -10.1188572
201 -34098.8161 -32242.61217 -30650.84348 -13919.31411 -10.3386138
200 -34459.78825 -18236.33859 -22702.95051 -14459.97419 -10.448059
199 -35026.55425 -1970.48827 -13092.20674 -16653.13992 -10.6199
198 -35388.06574 17163.48277 -2215.214565 -18000.07298 -10.7295087
197 -33849.40909 37540.4053 10673.37781 -17298.23548 -10.2629947
196 -30571.06317 58117.20125 24717.06855 -16545.93831 -9.26901436
195 -27596.03702 74787.10459 35724.72715 -15903.38629 -8.36699928
194 -24726.53922 83544.10667 40434.49523 -14485.33589 -7.49697993
193 -19899.3563 88564.42278 42446.86339 -12893.95927 -6.03339891
192 -14118.39137 92128.35875 46855.29191 -11873.9379 -4.28063531
191 -13080.66178 93202.74976 50516.52102 -12820.89153 -3.96600018
190 -18840.22752 95842.13563 50158.13368 -16132.2004 -5.71227564
 delta epsilon

B C D

-1.94117172 -2.05122664 -0.10857694 DELTA EPSILON IS UNIT OF ABSORBANCE

-2.0043313 -2.06551236 -0.16433304 WHATS THE POINT
-2.01071752 -2.06931431 -0.13199954
-2.02074932 -2.09362665 -0.08352748 det 2" struc
-2.09769727 -2.14186551 -0.05231666 know best denaturant
-2.21481379 -2.18145363 -0.03614618 get delta E
-2.34676059 -2.2250625 -0.07947028 use it to get %2" structure
-2.46609836 -2.32602773 -0.11669085 search K2D3. click the first thing
-2.57170356 -2.47414053 -0.11146219 wavelength: 190 to 250 nm
-2.77118516 -2.61794419 -0.1531324 CD Units delta epsilon
-3.05962213 -2.79581433 -0.19676544 protein size leave blank
-3.32795806 -3.04234917 -0.18661523 CD spectrum: delta epsilon
-3.67659134 -3.28237582 -0.19181051 * higher percent 2" less den
-4.18584665 -3.52466893 -0.20501627 trying to monitor conformation thru visc
-4.73444889 -3.86039798 -0.2133555
-5.30008919 -4.26603372 -0.26031929 * may include figure in discussion, but p
-5.92341446 -4.68537724 -0.3063307
-6.60809066 -5.13494634 -0.33436628
-7.39603396 -5.65232833 -0.4099971
-8.21274336 -6.20884045 -0.48199556
-8.97780428 -6.70465426 -0.46431864
-9.79259311 -7.14780895 -0.45773342
-10.6130885 -7.6483436 -0.49028123
-11.2874115 -8.1901398 -0.44955704
-11.9225974 -8.64291313 -0.47039306
-12.5873055 -8.95322684 -0.62331763
-13.1244588 -9.21415803 -0.61654989
-13.4961816 -9.4850148 -0.4641652
-13.793038 -9.67749293 -0.50422567
-14.0824653 -9.78440097 -0.65546322
-14.3438134 -9.8801186 -0.61106999
-14.4450894 -9.94681776 -0.50117048
-14.4180115 -9.99345184 -0.62538144
-14.4542752 -10.0285808 -0.78528061
-14.5605987 -10.0637714 -0.69878169
-14.69481 -10.2243382 -0.60858232
-14.9321537 -10.4796143 -0.68899568
-15.2382405 -10.7341606 -0.85761955
-15.6412887 -11.1142632 -1.11754102
-16.4164647 -11.6811494 -1.18687307
-17.5422553 -12.3819293 -0.9889402
-18.7045405 -13.1700416 -1.12965063
-19.7448657 -13.9469267 -1.63753384
-20.3752152 -14.5858652 -1.95861733
-20.3740835 -14.8372337 -2.20751483
-19.7907888 -14.6097431 -2.65068935
-18.4665522 -14.0133882 -3.15346408
-16.3548241 -12.8806405 -3.70993452
-13.4783588 -11.2304194 -4.15993646
-9.7758208 -9.29320341 -4.22027594
-5.52917912 -6.88343657 -4.38420174
-0.59744354 -3.96950056 -5.04916012
5.20389387 -0.67164349 -5.45754441
11.3820888 3.23612207 -5.24475031
17.6208845 7.49410847 -5.01665706
22.6751272 10.831583 -4.82183806
25.3302124 12.2595644 -4.3918913
26.8523506 12.8697057 -3.90939278
27.9329206 14.2063222 -3.60012671
28.2586713 15.3163911 -3.88723896
29.0589217 15.2077296 -4.89121351
 UNIT OF ABSORBANCE????????

k the first thing

elength: 190 to 250 nm
Units delta epsilon
ein size leave blank
pectrum: delta epsilon values
er percent 2" less denatured. best denaturant has low helicity
conformation thru viscosity and CD spectroscopy

re in discussion, but put in appendix

determine whether the denaturant is good or not
highest viscosity = best denaturant
a good denaturant must not affect the primary structure. should only affect H-bonds, but NOT

CD is like UV. may max absorption. we can correlate absorption to type of 2" structure.
radiate light; goes through filter. may be circular polarization -- circular dichroism
polarization of beta sheet is different from that of alpha helices

absorbance tells you about the 2" structure

alpha helix has a particular range of values; so does beta sheet
ffect H-bonds, but NOT THE PEPTIDE BONDS

of 2" structure.
dichroism