a
Sir3
Sir3

Sir3
Sir3

Sir3
Sir3

Sir3

Sir3
streptavidin bead capture
Sir3 Western

WT
[NaCl]:

H4K16Q

20mM

40mM

50mM

75mM

U

U

U

U

20mM

40mM

50mM

U

U

75mM

αSir3
B

B

B

B

U

B

B

B

U

B

Bound Sir3 (nM)

b
WT
H4K16Q

Total Sir3 (nM)

c

2.5 mM Na+
WT

40 mM Na+
H4K16Q

WT

H4K16Q

Sir3/N:

e

WT
0 Sir3/N
1 Sir3/N
2 Sir3/N
4 Sir3/N

Sedimentation coefficient (S)

Boundary fraction (%)

Boundary fraction (%)

d

H4K16Q
0 Sir3/N
1 Sir3/N
2 Sir3/N
4 Sir3/N

Sedimentation coefficient (S)

b

v (mL/g)

Boundary fraction (%)

a

Sedimentation coefficient (S)

c

R 2 = 0.933

slope = 0.00192
y-intercept = 0.627

Sedimentation coefficient (S)

Sed. Coefficient (s-13)

v (mL/g)

Mol. Weight (MDa)

Frictional Ratio

13.517

0.654

1.8714 (1.8307, 1.9122)

5.1595 (5.0840, 5.2350)

16.737

0.658

2.0831 (2.0147, 2.1515)

5.2967 (5.1809, 5.4125)

22.444

0.669

2.2274 (2.2223, 2.2326)

4.2431 (4.2389, 4.2474)

22.753

0.677

2.2628 (2.2313, 2.2944)

3.7372 (3.7018, 3.7727)

26.670

0.671

2.2293 (2.1968, 2.2619)

3.0956 (3.0651, 3.1262)

35.043

0.695

2.5851 [2.6144] (2.4896, 2.6806)

2.4714 (2.4077, 2.5351)

a

Molecular weight (Da)

Frictional ratio (f/f0)

Frictional ratio (f/f0)

b

Partial Concentration

Sedimentation coefficient (S)

H4K16Q + Sir3

Partial Concentration

Boundary fraction (%)

WT + Sir3

Molecular weight (Da)

v (mL/g)

Mol. Weight (MDa)

Sir3

Frictional Ratio

WT

0.694

2.5663 [2.6144] (2.5526, 2.5801)

N/A

2.2302 (2.2223, 2.2381)

H4K16Q

0.697

2.6667 [2.6144] (2.6356, 2.6978)

N/A

2.1616 (2.1446, 2.1787)

WT + Sir3

0.715

5.2706 (5.2297, 5.3115)

23.9

2.5824 (2.5694, 2.5953)

H4K16Q + Sir3

0.702

2.5380 (2.4936, 2.5824)

N/A

1.9981 (1.9746, 2.0215)

Frictional ratio (f/f0)

Frictional ratio (f/f0)

Folded

Molecular weight (Da)

Phosphate

Extended

Folded

c

Molecular weight (Da)

Partial Concentration

Sedimentation coefficient (s)

Frictional ratio (f/f0)

Molecular weight (Da)

Frictional ratio (f/f0)

Sedimentation coefficient (S)

Partial Concentration

Boundary fraction (%)

Extended

Partial Concentration

b

Tris

Partial Concentration

Boundary fraction (%)

a

Molecular weight (Da)

v (mL/g)

Mol. Weight (MDa)

Frictional Ratio

Tris

0.775

2.5109 [2.6144] (2.4730, 2.5487)

1.4925 (1.4778, 1.5073)

Tris + MgCl 2

0.576

2.4943 [2.6144] (2.4536, 2.5351)

2.5842 (2.5547, 2.6137)

Phosphate

0.699

2.6163 [2.6144] (2.4995, 2.7330)

2.0966 (2.0341, 2.1591)

Phosphate + MgCl 2

0.595

2.4558 [2.6144] (2.4378, 2.4737)

2.5157 (2.5032, 2.5281)

0.2 μm

0.2 μm

Extended (Tris)

Folded

Count

Height = 5.62 (5.14, 6.11)

Count

Height = 1.91 (1.85, 1.97)

0.2 μm

0.2 μm

Height (nm)
WT (Phosphate)

H4K16Q

Height = 1.84 (1.78, 1.90)

Count

Count

Height = 1.60 (1.56, 1.64)

0.2 μm
μm
0.2

0.2 μm

Height (nm)

Height (nm)
WT + Sir3

H4K16Q + Sir3

Height = 1.72 (1.64, 1.81)

Count

Count

Height = 2.79 (2.53, 3.05)

0.2 μm

0.2 μm

Height (nm)

Height (nm)

e

Folded

μm

WT

H4K16Q

WT + Sir3

WT + Sir3

0.1 μm

0.1 μm

0.1 μm

0.1 μm

0.1 μm

2

μm

3.7 nm

0.

2

μm

μm

0.

μm

5.0 nm

2

μm

H4K16Q + Sir3

0.1 μm

0.

2

μm

0.

2

μm

H4K16Q + Sir3

4.4 nm

0.

2

μm

0.

2

μm

2.5 nm

0.

2

2

0.

μm

2.0 nm

0.1 μm

μm

2

0.
2

0.

0.1 μm

μm

μm

2

2

0.

f

0.

0.

0.1 μm

Sir3

13 nm

3.8 nm

2

Extended

0.

d

3.6 nm

0.

2

μm

μm

c

2

b

Height (nm)

0.

a

Swygert et al., Fig. 6
WT + BAH

H4K16Q + BAH

Height  =  3.05  (2.67,  3.44)

Height  =  1.62  (1.54,  1.70)

Count

Count

a

0.2  μm

Height  (nm)

WT + BAH

WT + BAH

Q + BAH

Q + BAH

0.1  μm

0.1  μm

0.1  μm

0.1  μm

4.7nm

20  nm

2.8nm

0.

2  

μm

0.

2  
μm

2  
μm

μm

0.1  μm

0.

2  

2  
μm

0.

c

2.7nm

0.

μm

2  
μm

2  

0.

0.

2  
μm

2.7nm

0.

b

0.2  μm

Height  (nm)

50  nm

BAH

0.1  μm

a

b

c

Array in Tris

Array in Phosphate

Array in Magnesium

Array + Sir3

Sir3

BAH
C

d

Array + Sir3 BAH