Structural insights into proteoglycan-shaped Hedgehog signaling

Whalen, Daniel M.; Malinauskas, Tomas; Gilbert, Robert J.C.; Siebold, Christian

doi:10.1073/pnas.1310097110

Cited by 91 publications

(91 citation statements)

References 43 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Deciphering the exact 3D organization and features of BMP dimers will require further analysis, and the contribution of single or groups of basic residues -distant from the CW motifs-will need to be taken into consideration to precisely delineate the overall structural basis of BMP-HS interactions (25,46,47). It will be necessary also to clarify the spatial orientation of different BMPs with respect to the HS chain backbone given that their electropositive surfaces differ in location and structure as suggested above.…”

Section: Discussionmentioning

confidence: 99%

Domains with highest heparan sulfate–binding affinity reside at opposite ends in BMP2/4 versus BMP5/6/7: Implications for function

Billings

Yang

Mundy

et al. 2018

Journal of Biological Chemistry

View full text Add to dashboard Cite

Signaling proteins, including bone morphogenetic proteins (BMPs), specifically interact with heparan sulfate (HS). These interactions regulate protein distribution and function and are largely mediated by domains rich in basic amino acids. The N-terminal region of BMP2 and BMP4 contains one such domain with a typical Cardin-Weintraub (CW) motif, but it is unclear whether the same occurs in BMP5, BMP6 and BMP7 that constitute a separate evolutionary subgroup. Peptides spanning the N-terminal domain of BMP2/4 interacted with substrate-bound HS with nanomolar affinity, but peptides spanning BMP5/6/7 N-terminal domain did not. We reexamined the entire BMP5/6/7 sequences and identified a novel CW-like motif at their Cterminus. Peptides spanning this domain displayed high affinity HS binding, but corresponding BMP2/4 C-terminal peptides did not, likely because of acidic or non-charged residue substitutions. Peptides pre-assembled into NeutrAvidin tetramers displayed the same exact binding selectivity of respective monomers, but bound HS with greater affinity. Tests of possible peptide biological activities showed that the HS-binding N-terminal BMP2/4 and C-terminal BMP5/6/7 peptides stimulated chondrogenesis in vitro, potentially by freeing endogenous BMPs. Thus, HS interactions appear largely ascribable to domains at opposite ends of BMP2/4 versus BMP5/6/7, reiterating the evolutionary distance of these BMP subgroups and possible functional diversification.

show abstract

Section: Discussionmentioning

confidence: 99%

Domains with highest heparan sulfate–binding affinity reside at opposite ends in BMP2/4 versus BMP5/6/7: Implications for function

Billings

Yang

Mundy

et al. 2018

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…Heparin 15-mer required to induce dimerization; orientation of GAG chains in complex depends on sulfation pattern (Whalen et al, 2013) Heparin binding: adhesion molecules Integrins (avb3) Molecular modeling (docking) A conserved heparin binding site identified in RGD integrins but not in non-RGD integrins (Ballut et al, 2013) P-selectin, L-selectin ELISA assays, cell adhesion assays Hexasulfated tetrasaccharide (Nelson et al, 1993) (continued ) 80 Octasaccharide and decasaccharide bind thrombospondin similarly to synthetic pentasaccharide (Tan et al, 2006) and induce formation of trans-and cis-dimers respectively (Tan et al, 2008) Complement system and innate immunity Factor H Analytical ultracentrifugation, X-ray scattering, SPR Cooperative, bivalent binding to heparin of long oligosaccharides (dp18 and over) (Khan et al, 2012) Affinity chromatography, microtiter plate binding assays Two GAG binding sites on factor H select different sulfation patterns (Clark et al, 2013) Human b-defensin 2 Gel mobility shift, NMR titration with synthetic heparin pentasaccharide (and DS hexasaccharide)…”

Section: Sonic Hedgehogmentioning

confidence: 99%

Pharmacology of Heparin and Related Drugs

et al. 2015

View full text Add to dashboard Cite

“…The palmitoylated N-terminal Shh peptide includes the CardinWeintraub motif (27), a short stretch rich in positively charged residues (K32-K38 in human Shh), which contributes to Shh binding to glycosaminoglycans (GAGs) (28). It is conceivable that the P26 mutation might affect Shh interaction with GAGs.…”

Section: Holoprosencephaly-causing Shh Mutation Abolishes Palmitate-dmentioning

confidence: 99%

Mechanism of inhibition of the tumor suppressor Patched by Sonic Hedgehog

Tukachinsky

Petrov

Watanabe

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

The Hedgehog cell–cell signaling pathway is crucial for animal development, and its misregulation is implicated in numerous birth defects and cancers. In unstimulated cells, pathway activity is inhibited by the tumor suppressor membrane protein, Patched. Hedgehog signaling is triggered by the secreted Hedgehog ligand, which binds and inhibits Patched, thus setting in motion the downstream events in signal transduction. Despite its critical importance, the mechanism by which Hedgehog antagonizes Patched has remained unknown. Here, we show that vertebrate Patched1 inhibition is caused by direct, palmitate-dependent interaction with the Sonic Hedgehog ligand. We find that a short palmitoylated N-terminal fragment of Sonic Hedgehog binds Patched1 and, strikingly, is sufficient to inhibit it and to activate signaling. The rest of Sonic Hedgehog confers high-affinity Patched1 binding and internalization through a distinct binding site, but, surprisingly, it is not absolutely required for signaling. The palmitate-dependent interaction with Patched1 is specifically impaired in a Sonic Hedgehog mutant causing human holoprosencephaly, the most frequent congenital brain malformation, explaining its drastically reduced potency. The palmitate-dependent interaction is also abolished in constitutively inhibited Patched1 point mutants causing the Gorlin cancer syndrome, suggesting that they might adopt a conformation distinct from the wild type. Our data demonstrate that Sonic Hedgehog signals via the palmitate-dependent arm of a two-pronged contact with Patched1. Furthermore, our results suggest that, during Hedgehog signaling, ligand binding inhibits Patched by trapping it in an inactive conformation, a mechanism that explains the dramatically reduced activity of oncogenic Patched1 mutants.

show abstract

Structural insights into proteoglycan-shaped Hedgehog signaling

Cited by 91 publications

References 43 publications

Domains with highest heparan sulfate–binding affinity reside at opposite ends in BMP2/4 versus BMP5/6/7: Implications for function

Domains with highest heparan sulfate–binding affinity reside at opposite ends in BMP2/4 versus BMP5/6/7: Implications for function

Pharmacology of Heparin and Related Drugs

Mechanism of inhibition of the tumor suppressor Patched by Sonic Hedgehog

Contact Info

Product

Resources

About