An Emerging Role of Sonic Hedgehog Shedding as a Modulator of Heparan Sulfate Interactions

Ohlig, Stefanie; Pickhinke, Ute; Sirko, Swetlana; Bandari, Shyam; Hoffmann, Daniel; Dreier, Rita; Farshi, Pershang; Götz, Magdalena; Grobe, Kay

doi:10.1074/jbc.m112.356667

Cited by 56 publications

(49 citation statements)

References 72 publications

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“…B 369: 20130545 shifted protein. Consistent with these findings, in mammalian cell cultures, the release of soluble and oligomerized Shh from the membrane is considered to be mediated by HS-dependent mechanisms [78][79][80]. These data suggest that HSPGs can influence lipid-linked growth factor signalling-at least in part-by binding to lipoproteins.…”

Section: Divergent Functions Of Heparan Sulfatesupporting

confidence: 57%

Extracellular distribution of diffusible growth factors controlled by heparan sulfate proteoglycans during mammalian embryogenesis

Matsuo

Kimura-Yoshida

2014

Phil. Trans. R. Soc. B

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During mouse embryogenesis, diffusible growth factors, i.e. fibroblast growth factors, Wnt, bone morphogenetic protein and Hedgehog family members, emanating from localized areas can travel through the extracellular space and reach their target cells to specify the cell fate and form tissue architectures in coordination. However, the mechanisms by which these growth factors travel great distances to their target cells and control the signalling activity as morphogens remain an enigma. Recent studies in mice and other model animals have revealed that heparan sulfate proteoglycans (HSPGs) located on the cell surface (e.g. syndecans and glypicans) and in the extracellular matrix (ECM; e.g. perlecan and agrin) play crucial roles in the extracellular distribution of growth factors. Principally, the function of HSPGs depends primarily on the fine features and localization of their heparan sulfate glycosaminoglycan chains. Cellsurface-tethered HSPGs retain growth factors as co-receptors and/or endocytosis mediators, and enzymatic release of HSPGs from the cell membrane allows HSPGs to transport or move multiple growth factors. By contrast, ECM-associated HSPGs function as a reservoir or barrier in a context-dependent manner. This review is focused on our current understanding of the extracellular distribution of multiple growth factors controlled by HSPGs in mammalian development.

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Section: Divergent Functions Of Heparan Sulfatesupporting

confidence: 57%

Extracellular distribution of diffusible growth factors controlled by heparan sulfate proteoglycans during mammalian embryogenesis

Matsuo

Kimura-Yoshida

2014

Phil. Trans. R. Soc. B

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“…This led us to propose that inactive SHH is transported, requiring an activation step after long-range transport (Feng et al, 2004). More recently, Zn 2+ mediated metalloprotease cleavage by ADAM (Ohlig et al, 2012), resulting in N-terminal lipid removal, was proposed to be required for SHH activation. However, neither report was able to show that SHH activation occurs through multimer disassembly or cleavage events, in vivo, or to determine in vivo sites where such activation occurs.…”

Section: Discussionmentioning

confidence: 99%

SHH E176/E177-Zn2+ conformation is required for signaling at endogenous sites

Himmelstein

Cajigas

et al. 2017

Developmental Biology

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Summary Sonic hedgehog (SHH) is a master developmental regulator. In 1995, the SHH crystal structure predicted that SHH-E176 (human)/E177 (mouse) regulates signaling through a Zn2+-dependent mechanism. While Zn2+ is known to be required for SHH protein stability, a regulatory role for SHH-E176 or Zn2+ has not been described. Here, we show that SHH-E176/177 modulates Zn2+-dependent cross-linking in vitro and is required for endogenous signaling, in vivo. While ectopically expressed SHH-E176A is highly active, mice expressing SHH-E177A at endogenous sites (ShhE177/−) are morphologically indistinguishable from mice lacking SHH (Shh−/−), with patterning defects in both embryonic spinal cord and forebrain. SHH-E177A distribution along the embryonic spinal cord ventricle is unaltered, suggesting that E177 does not control long-range transport. While SHH-E177A association with cilia basal bodies increases in embryonic ventral spinal cord, diffusely distributed SHH-E177A is not detected. Together, these results reveal a novel role for E177-Zn2+ in regulating SHH signaling that may involve critical, cilia basal-body localized changes in cross-linking and/or conformation.

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“…The effect of the N-terminal Bzm modification on heparin binding was assessed by heparin affinity-chromatography [55, 56, 58]. Unmodified ShhN, NEM-ShhN and Bzm-ShhN had comparable binding to heparin, with all forms eluting at ~1 M NaCl (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The conserved N-terminus of ShhN extends away from the core structure [58] and provides a somewhat unstructured and flexible arm to probe for binding interactions in close proximity. When Bzm-ShhN was UV irradiated alone, no crosslinked products were observed by SDS-PAGE/Western blotting.…”

Section: Discussionmentioning

confidence: 99%

Design and characterization of a photo-activatable hedgehog probe that mimics the natural lipidated form

House¹,

Daye²,

Tarpley³

et al. 2015

Archives of Biochemistry and Biophysics

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We have generated a photoactivatable form of sonic hedgehog protein by modifying the N-terminal cysteine with the heterobifunctional photocrosslinker 4-maleimidobenzophenone (Bzm). The Bzm modification on ShhN imparted a significant increase in activity as assessed in the C3H10T1/2 functional assay with potency comparable to that of the endogenous dual-lipidated form of ShhN (ShhNp). Reversed-phase HPLC analysis indicated that the increase in activity compared to unmodified ShhN may be due in part to the hydrophobic nature of the benzophenone group. In contrast to the fully processed ShhNp, Bzm-ShhN is monomeric as assessed by analytical SEC and does not require detergent to be soluble. Further, we demonstrated that the Bzm-ShhN was able to crosslink in vitro in the presence of a known binding partner, heparin. We suggest that Bzm-ShhN can serve as a relatively facile and preferred source of ShhNp for in vitro assays and as a probe to identify novel Hh protein interactions.

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An Emerging Role of Sonic Hedgehog Shedding as a Modulator of Heparan Sulfate Interactions

Cited by 56 publications

References 72 publications

Extracellular distribution of diffusible growth factors controlled by heparan sulfate proteoglycans during mammalian embryogenesis

Extracellular distribution of diffusible growth factors controlled by heparan sulfate proteoglycans during mammalian embryogenesis

SHH E176/E177-Zn2+ conformation is required for signaling at endogenous sites

Design and characterization of a photo-activatable hedgehog probe that mimics the natural lipidated form

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