| RRC ID |
39838
|
| Author |
Saeki K, Katsuura M, Yanagisawa S, Suzuki R, Okazaki M, Kimura M.
|
| Title |
Inactivation of N-terminal signaling domain of Sonic hedgehog by forming a disulfide bond.
|
| Journal |
Biochim Biophys Acta
|
| Abstract |
The N-terminal domain of mouse Sonic hedgehog (Shh-N) expressed in mammalian cells showed four-fold bands on non-reduced SDS-PAGE, though it was homogeneous under reduced conditions. It contains three cysteine residues, Cys-25, Cys-103, and Cys-184, which may be concerned with this heterogeneity. Therefore, we examined the formation of a disulfide bond in the recombinant Shh-N and identified three kinds of disulfides with a combination of peptide mapping and NH(2)-terminal amino acid sequencing analysis. Among them, one type of the Shh-N containing a disulfide bond of Cys-103/Cys-184 could be separated from the other Shh-Ns using reverse phase HPLC and had no activity of alkaline phosphatase induction in C3H10T1/2 cells. This molecule could also be made by denaturation of the purified Shh-N with guanidine-HCl under non-reduced conditions. On the other hand, the reduced Shh-N and the reduced S-methylated Shh-N at cysteine residues showed approximately 10-fold higher activity compared to the originally purified Shh-N. These results suggested that Shh-N was synthesized as an active form whose three cysteine residues did not form disulfide and inactivated finally by forming a disulfide bond between Cys-103 and Cys-184.
|
| Volume |
1476(2)
|
| Pages |
219-29
|
| Published |
2000-2-9
|
| DOI |
10.1016/s0167-4838(99)00254-x
|
| PII |
S0167-4838(99)00254-X
|
| PMID |
10669787
|
| MeSH |
Animals
Disulfides
Hedgehog Proteins
Mice
Protein Conformation
Proteins / chemistry*
Proteins / metabolism
Recombinant Proteins / chemistry
Recombinant Proteins / metabolism
Trans-Activators*
|
| IF |
3.411
|
| Times Cited |
2
|
|
WOS Category
|
BIOPHYSICS
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| Resource |
| Human and Animal Cells |
L cells |
