folding of thermophilic c-domain i m r g = 46 Å n r g = 38 Å i m state never populates i m state...

118

Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å • I I m state never state never populates populates I t R g = 60 Å • On pathway folding On pathway folding P15.1 G U C U U c A u G c U C GG G U A A U C G A A A AC C C A AA U g a U G G U A G G G G c A C C U U C c c g A a G G A A a U g A A C G G A G G G A A G G A C A G g c g c a u g c C U G U A U A G A U G A U U a C CGC C g G AG U A C G A G G c G c a a A G C C G c U U G C A G U A C G A a G G u A C A g A A C A U G G C U U A u A G A g C a U g A G A C C A C U U G U C C U G C U C G C A C G G U G C U G A G A U G C C C G U A G U GU GCGAG C U G C g G c c u u u g gg C c GUAG A G G A A A 1 20 40 60 260 280 300 320 340 360 380 250 80 P1 P2 P4 P15 P18 P19 P3 P5 P5.1 5' 3' G U U C C A G A A g c g c a g 400 420 g c A G L1

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Page 1: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Folding of thermophilic C-domain

Im Rg = 46 Å

N Rg = 38 Å

• IImm state never populates state never populates

It Rg = 60 Å

• On pathway foldingOn pathway foldingP15.1

GU

C

U

U c A u G c U C G GGUAAUC

G

A

A A A C C CA A A U

gaUGGUA

GGGGcAC CU U C c c

gA a G GA

AaUg

AACG

GAGGGAAGG

ACAGg cgcau

gcC

UGU

AUAGAUGAUUaCC GC

CgGA G

UACGA G

GcGc a

aAG

CC G

cUU

GCAGUACGAaG

G u A C AgAACAUGGCUU

A

uAGAgCaUgAGA

CCACUU

GUCC

UGCUCGC

ACGGU

GCUG

AGAUGCCCGU

AGUG U

G C G A G

CUGCgGccuuug g g C c GU AG

AGGAAA

1 20

40

60

260

280

300

320

340

360

380

250

80

P1P2

P4

P15

P18

P19

P3

P5

P5.1

5'

3'

G

U

U

C

C

AG

A

A

g c

gca

g

400

420

g

c

AG

L1

B. stearothermophilus

Page 2: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Rg = 60 Å

Page 3: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 4: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 5: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 6: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 7: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 8: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 9: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 10: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 11: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 12: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 13: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 14: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 15: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 16: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 17: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 18: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 19: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 20: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 21: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 22: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 23: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 24: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 25: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 26: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 27: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 28: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 29: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 30: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 31: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 32: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 33: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 34: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 35: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 36: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 37: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 38: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 39: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

MgMg2+2+ conc. reaches K conc. reaches KMgMg((IImm-to--to-NN) ) firstfirst

Rg = 60 Å

99.98%

0.01% 0.01%

Neither Neither IImm nor nor NN populates yet populates yet

Page 40: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 41: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 42: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 43: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 44: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 45: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 46: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 47: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 48: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 49: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 50: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 51: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 52: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 53: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 54: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 55: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 56: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 57: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 58: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 59: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 60: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 61: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 62: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 63: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 64: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 65: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 66: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 67: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 68: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 69: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 70: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 71: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

0.1%

Rg = 60 Å

Rg = 46 Å

Rg = 38 Å

10%90%

N starts accumulating, It still dominantsIItt is the is the reference statereference state for the stability of for the stability of NNIImm is is not importantnot important for the stability of for the stability of NN

Page 72: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 73: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 74: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 75: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 76: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 77: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Rg = 60 ÅRg = 38 Å

50%

MgMg2+2+ conc. reaches conc. reaches KKMgMg(functional)(functional)

49.5%

~ KMg(It-to-N)

0.5%

Im is negligible

Page 78: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 79: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 80: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 81: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 82: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 83: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 84: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 85: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 86: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 87: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 88: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 89: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 90: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 91: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Rg = 60 Å

Rg = 38 Å

96%

N-stateN-state dominants dominants

3.4%

IItt is the reference state is the reference state

0.6%

Im is negligible

Page 92: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 93: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 94: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 95: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 96: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 97: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 98: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 99: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 100: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Rg = 38 Å

99.2%

0.4% 0.4%

When MgWhen Mg2+2+ reaches K reaches KMgMg(I(Itt-to-I-to-Imm), ),

N-stateN-state is over 99% is over 99%

Page 101: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 102: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 103: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 104: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 105: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 106: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 107: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 108: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 109: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 110: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 111: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 112: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 113: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 114: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 115: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 116: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 117: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Page 118: Folding of thermophilic C-domain I m R g = 46 Å N R g = 38 Å I m state never populates I m state never populates I t R g = 60 Å On pathway folding On

Rg = 38 Å

99.91%

Rg = 60 Å Rg = 46 Å

0.08%0.006%

At high enough Mg2+, Im becomes more stable than It

But none of them populates

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