|
Sample contents
|
Remaining % of NCS-Chra
|
|---|
|
NCS-Chr
|
0%
|
|
NCS-Chr + 1.2 M GdnHCl
|
13 ± 4%b
|
|
holoNCS (NCS-Chr + 1:1 folded state of apoNCS)
|
100 ± 4%
|
|
NCS-Chr + 1:1 MG-like state of apoNCS (in 1.2 M GdnHCl)
|
101 ± 4%c
|
- aSamples containing 20 μM NCS-Chr were incubated with folded or MG-like state of apoNCS in 10 mM phosphate aqueous buffer at pH 3 and 25°C for 30 min. Remaining amount of intact NCS-Chr was analyzed by HPLC.
- bMost NCS-Chr was degraded in acidic aqueous condition in the presence of 1.2 M GdnHCl. The small remaining amount of the NCS-Chr exhibited additional forms. About 35 ± 10% of the remaining NCS-Chr was converted into chlorohydrin form of the chromophore. It is known that the chlorohydrin derivative of NCS-Chr can be obtained by treatment of HCl [2]. Presumably, excess amount of GdnHCl triggers the conversion process.
- cNo chlorohydrin conversion could be found in the remaining NCS-Chr in the presence of MG-like state of apoNCS. Presumably, binding of apoNCS prevents the chromophore conversion reaction triggered by excess amount of GdnHCl.
