Bcakground

Sonic Hedgehog (Shh) is expressed in embryonic tissues that are critical for the patterning of the developing central nervous system, somite, and limb. It is also involved in whisker, hair, foregut, tooth, and bone development. Shh regulates neural and hematopoietic stem cell fate and is important for thymocyte differentiation and proliferation as well as T cell determination. In adult tissue Shh is associated with cancer development and tissue remodeling following injury (1-3). Human Shh encodes a 462 amino acid (aa) precursor protein that is autocatalytically processed to yield a non-glycosylated 19 kDa N-terminal fragment (Shh-N) and a glycosylated 25 kDa C-terminal protein (Shh-C) (4). Shh-C, which is responsible for the intramolecular processing of Shh, is rapidly degraded following Shh proteolysis (5). Shh-N is highly conserved, sharing >98% aa identity between mouse, human, rat, canine, porcine, and chicken Shh-N. Shh-N can be palmitoylated at its N-terminal cysteine and modified by cholesterol addition at its C-terminus (6). These modifications contribute to the membrane tethering of Shh as well as its assembly into various sized multimers (6-9). Lipid modification and multimerization greatly increase Shh-N receptor binding affinity and signaling potency (5, 6, 8, 9). Monomeric and multimeric Shh can be released from the plasma membrane by the cooperative action of DISP1, SCUBE2, and TACE/ ADAM17 (10-12). Modifications also extend the effective range of Shh functionality and are required for the development of protein gradients important in tissue morphogenesis (9, 13). Canonical signaling of Shh is mediated by a multicomponent receptor complex that includes Patched (PTCH1, PTCH2) and Smoothened (SMO) (14)

Reference

1. Briscoe, J. and P.P. Therond (2013) Mol. Cell. Biol. 14:416.

2. Aviles, E.C. et al. (2013) Front. Cell. Neurosci. 7:86.

3. Xie, J. et al. (2013) OncoTargets Ther. 6:1425.

4. Marigo, V. et al. (1995) Genomics 28:44.

5. Zeng, X. et al. (2001) Nature 411:716.

6. Feng, J. et al. (2004) Development 131:4357.

7. Goetz, J.A. et al. (2006) J. Biol. Chem. 281:4087.

8. Pepinsky, R.B. et al. (1998) J. Biol. Chem. 273:14037.

9. Pepinsky, R.B. et al. (1998) J. Biol. Chem. 273:14037.

10. Chen, M.-H. et al. (2004) Genes Dev. 18:641.

11. Etheridge, L.A. et al. (2010) Development 137:133.

12. Jakobs, P. et al. (2014) J. Cell Sci. 127:1726.

13. Dierker, T. et al. (2009) J. Biol. Chem. 284:8013.

14. Lewis, P.M. et al. (2001) Cell 105:599.

Specifications

Synonyms	Shh;HHG1; HHG-1; HLP3; HPE3; MCOPCB5;Sonic Hedgehog; TPT; TPTPS
Accession #	Q15465
Source	Human embryonic kidney cell, HEK293-derived human Sonic Hedgehog/Shh protein
Source	Cys24-Gly197
Predicted Moleucular weight	19.6 kDa
Form/Structure	Dimer in solution

Components and Storage

Formulation	Solution protein
	Dissolved in sterile PBS buffer
	This solution can be diluted into other aqueous buffers. Centrifuge the vial prior to opening
Storage and Stability	Avoid repeated freeze-thaw cycles
	It is recommended that the protein be aliquoted for optimal storage
	12 months from date of receipt, -20 to -70 °C as supplied
Shipping	Shipping with dry ice

Quality

Purity	> 95%, determined by SDS-PAGE.
Endotoxin Level	<0.010 EU per 1 ug of the protein by the LAL method.
Activity	Measured by its ability to induce alkaline phosphatase production by C3H10T1/2 mouse embryonic fibroblast cells.
Activity	The EC50 for this effect is typically 5-15 ng/mL.

SDS-PAGE

Bioactivity

Documents

Human SHH