TABLE 2.

Hhat palmitoylation: reaction conditions and kinetics

Optimization of in vitro palmitoylation reaction conditions
	pH	[NaCl]	[DTT]	[Triton X-100]	[ATP]	[Divalent cation]
Initial	100 mm HEPES (pH 7.3)	200 mm	1 mm	0.05%	1 mm	1 mm MgCl2
Final	100 mm MES (pH 6.5)	20 mm	1 mm	0.05%	0	0
Required			Yes	Yes	No	No

Kinetic measurements
Shh titration at 100 μm [125I]iodopalmitoyl-CoA		[125I]Iodopalmitoyl-CoA titration at 50 μm Shh
Vmax	Km	Vmax	Km
pmol/min	μm	pmol/min	μm
0.25 ± 0.03	1.25 ± 0.26	0.21 ± 0.03	3.0 ± 0.28