Table 1.

Amino acid sequence, thermal unfolding midpoints (Tm), and apparent molecular weight of VHP subdomains

Protein	Sequence																																				Tm	Apparent MW
		42					47				51		53					58																		76
HP36	M	L	S	D	E	D	F	K	A	V	F	G	M	T	R	S	A	F	A	N	L	P	L	W	K	Q	Q	N	L	K	K	E	K	G	L	F	70	3100
M53L							|				|		L					|																			62	3100
F47L							L				|		L					|																			45	3400
F51L							|				L		L					|																			50	3400
F58L							|				|		L					L																			35	3400
F47, 51L							L				L		L					|																			45	4100
F47, 58L							L				|		L					L																			(39)	7400
F51, 58L							|				L		L					L																			(37)	7700
HP35		L	S	D	E	D	F	K	A	V	F	G	M	T	R	S	A	F	A	N	L	P	L	W	K	Q	Q	N	L	K	K	E	K	G	L	F
		1					6				10		12					17																		35

Only the changed residues are shown for the mutant sequences. All of the leucine mutants also contain the M53L mutation. HP35 corresponds to last 35 residues (791 to 825) of intact chicken villin (Bazari et al. 1988). HP36 corresponds to the expressed VHP subdomain that retains an N-terminal methionine residue. The numbering at the top is relative to the 76-residue chicken villin headpiece, and corresponds to the numbering of the HP36 NMR structure 1VII.pdb (McKnight et al. 1997). The numbers at the bottom corresponds to the length of HP35. The T_m values were determined from the thermal unfolding experiments in Figure 2 ▶. The T_m values from the shallow unfolding curves of the double mutants that do not adopt the VHP subdomain native structure are in parentheses. The apparent molecular weights were calculated from the standards in the gel filtration experiments in Figure 5 ▶. The theoretical molecular weights are: HP36 = 4189.9; M53L = 4040.7; F47L, F51L, and F58L = 4006.6; F47, 51L, F47, 58L, and F51, 58L = 3972.6.