scholarly journals Structure and function of the ROR2 cysteine-rich domain in vertebrate noncanonical WNT5A signaling

2021 ◽  
Author(s):  
Samuel C Griffiths ◽  
Jia Tan ◽  
Armin Wagner ◽  
Levi L Blazer ◽  
Jarret J Adams ◽  
...  
Keyword(s):  
Receptor Tyrosine Kinase ◽  
Structure And Function ◽  
Hydrophobic Pocket ◽  
Rich Domain ◽  
Frizzled Receptors ◽  
Receptor Action ◽  
Robinow Syndrome ◽  
Modified Proteins ◽  
And Function ◽  
Cysteine Rich Domain

The receptor tyrosine kinase ROR2 mediates noncanonical WNT5A signaling to orchestrate tissue morphogenetic processes, and dysfunction of the pathway causes Robinow syndrome, Brachydactyly B and metastatic diseases. The domain(s) and mechanisms required for ROR2 function, however, remain unclear. We solved the crystal structure of the extracellular cysteine rich (CRD) and Kringle (Kr) domains of ROR2 and found that, unlike other CRDs, the ROR2 CRD lacks the signature hydrophobic pocket that binds lipids/lipid-modified proteins, such as WNTs, suggesting a novel mechanism of receptor action. Functionally, we showed that the ROR2 CRD, but not other domains, is required and minimally sufficient to promote WNT5A signaling, and Robinow mutations in the CRD and the adjacent Kr alter ROR2 function. Moreover, we demonstrated that the activity of the ROR2 CRD requires Frizzled receptors. Thus, ROR2 acts via its CRD to potentiate the function of a receptor supercomplex that includes Frizzleds to transduce WNT5A signals.

scholarly journals Probing Structure and Function of Alkali Sensor IRR with Monoclonal Antibodies

Biomolecules ◽  
2020 ◽  
Vol 10 (7) ◽  
pp. 1060
Author(s):  
Alexander S. Goryashchenko ◽  
Andrey A. Mozhaev ◽  
Oxana V. Serova ◽  
Tatiana N. Erokhina ◽  
Alexander N. Orsa ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Tyrosine Kinase ◽  
Insulin Receptor ◽  
Receptor Tyrosine Kinase ◽  
Specific Binding ◽  
Structure And Function ◽  
Full Length ◽  
Orphan Receptor ◽  
Extracellular Region ◽  
And Function

To study the structure and function of the pH-regulated receptor tyrosine kinase insulin receptor-related receptor (IRR), а member of the insulin receptor family, we obtained six mouse monoclonal antibodies against the recombinant IRR ectodomain. These antibodies were characterized in experiments with exogenously expressed full-length IRR by Western blotting, immunoprecipitation, and immunocytochemistry analyses. Utilizing a previously obtained set of IRR/IR chimeras with swapped small structural domains and point amino acid substitutions, we mapped the binding sites of the obtained antibodies in IRR. Five of them showed specific binding to different IRR domains in the extracellular region, while one failed to react with the full-length receptor. Unexpectedly, we found that 4D5 antibody can activate IRR at neutral pH, and 4C2 antibody can inhibit activation of IRR by alkali. Our study is the first description of the instruments of protein nature that can regulate activity of the orphan receptor IRR and confirms that alkali-induced activation is an intrinsic property of this receptor tyrosine kinase.

scholarly journals Ror2/Frizzled Complex Mediates Wnt5a-Induced AP-1 Activation by Regulating Dishevelled Polymerization

2010 ◽  
Vol 30 (14) ◽  
pp. 3610-3619 ◽  
Author(s):  
Michiru Nishita ◽  
Sumiyo Itsukushima ◽  
Akira Nomachi ◽  
Mitsuharu Endo ◽  
ZhiChao Wang ◽  
...  
Keyword(s):  
Tyrosine Kinase ◽  
Receptor Tyrosine Kinase ◽  
Wnt Pathway ◽  
Receptor Complex ◽  
Jnk Pathway ◽  
Rich Domain ◽  
Canonical Wnt Pathway ◽  
Receptor Component ◽  
Canonical Wnt ◽  
Cysteine Rich Domain

ABSTRACT The receptor tyrosine kinase Ror2 acts as a receptor or coreceptor for Wnt5a to mediate Wnt5a-induced activation of the Wnt/JNK pathway and inhibition of the β-catenin-dependent canonical Wnt pathway. However, little is known about how Ror2 cooperates with another receptor component(s) to mediate Wnt5a signaling. We show here that Ror2 regulates Wnt5a-induced polymerization of Dishevelled (Dvl) and that this Ror2-mediated regulation of Dvl is independent of the cytoplasmic region of Ror2. Ror2 can associate with Frizzled7 (Fz7) via its extracellular cysteine-rich domain to form a receptor complex that is required for the regulation of Dvl and activation of the AP-1 promoter after Wnt5a stimulation. Suppressed expression of Fz7 indeed results in the inhibition of Wnt5a-induced polymerization of Dvl and AP-1 activation. Interestingly, both the DIX and the DEP domains of Dvl are indispensable for Dvl polymerization and subsequent AP-1 activation after Wnt5a stimulation. We further show that polymerized Dvl is colocalized with Rac1 and that suppressed expression of Rac1 inhibits Wnt5a-induced AP-1 activation. Collectively, our results indicate that Ror2/Fz receptor complex plays an important role in the Wnt5a/Rac1/AP-1 pathway by regulating the polymerization of Dvl.

Mechanism and function of DHHC S-acyltransferases

2013 ◽  
Vol 41 (1) ◽  
pp. 29-34 ◽  
Author(s):  
Maurine E. Linder ◽  
Benjamin C. Jennings
Keyword(s):  
Fatty Acids ◽  
Protein S ◽  
Enzyme Mechanism ◽  
Long Chain Fatty Acids ◽  
Post Translational Modification ◽  
Rich Domain ◽  
Cytoplasmic Face ◽  
Protein Membrane ◽  
And Function ◽  
Cysteine Rich Domain

Protein S-palmitoylation is a reversible post-translational modification of proteins with fatty acids. In the last 5 years, improved proteomic methods have increased the number of proteins identified as substrates for palmitoylation from tens to hundreds. Palmitoylation regulates protein membrane interactions, activity, trafficking and stability and can be constitutive or regulated by signalling inputs. A family of PATs (protein acyltransferases) is responsible for modifying proteins with palmitate or other long-chain fatty acids on the cytoplasmic face of cellular membranes. PATs share a signature DHHC (Asp-His-His-Cys) cysteine-rich domain that is the catalytic centre of the enzyme. The biomedical importance of members of this family is underscored by their association with intellectual disability, Huntington's disease and cancer in humans, and raises the possibility of DHHC PATs as targets for therapeutic intervention. In the present paper, we discuss recent progress in understanding enzyme mechanism, regulation and substrate specificity.

Sprouty: a controversial role in receptor tyrosine kinase signalling pathways

2003 ◽  
Vol 31 (6) ◽  
pp. 1445-1446 ◽  
Author(s):  
X. Li ◽  
L. Wheldon ◽  
J.K. Heath
Keyword(s):  
Tyrosine Kinase ◽  
Receptor Tyrosine Kinase ◽  
Functional Divergence ◽  
Signalling Pathways ◽  
Signalling Molecules ◽  
Rich Domain ◽  
Growth Factor Signalling ◽  
Cysteine Rich Domain ◽  
Growth Factor Signalling Pathway

Sprouty was first identified in Drosophila as a novel antagonist of the fibroblast growth factor signalling pathway. Sprouty proteins comprise a big family, members of which are characterized by a cysteine-rich domain which confers inhibitory activity, whereas differences in the N-terminal region may be responsible for functional divergence. The role of Sprouty in RTK (receptor tyrosine kinase) signalling pathways is still controversial. Sprouty may negatively or positively regulate RTK signalling via differential interaction with different signalling molecules, and hence exert different mechanism of action.

scholarly journals Structure and function of NTAK, a novel ligand of receptor tyrosine kinase ErbB.

2000 ◽  
Vol 44 (2) ◽  
pp. 125-130
Author(s):  
Shigeki Higashiyama ◽  
Hiroshi Ishiguro ◽  
Toshiharu Nagatsu ◽  
Naoyuki Taniguchi
Keyword(s):  
Tyrosine Kinase ◽  
Receptor Tyrosine Kinase ◽  
Structure And Function ◽  
And Function

scholarly journals Structural Basis of Wnt Recognition by Frizzled

Science ◽  
2012 ◽  
Vol 337 (6090) ◽  
pp. 59-64 ◽  
Author(s):  
Claudia Y. Janda ◽  
Deepa Waghray ◽  
Aron M. Levin ◽  
Christoph Thomas ◽  
K. Christopher Garcia
Keyword(s):  
Binding Sites ◽  
Palmitoleic Acid ◽  
Index Finger ◽  
Cross Reactivity ◽  
Structural Basis ◽  
Hydrophobic Amino Acid ◽  
Rich Domain ◽  
Deep Groove ◽  
Frizzled Receptors ◽  
Cysteine Rich Domain

Wnts are lipid-modified morphogens that play critical roles in development principally through engagement of Frizzled receptors. The 3.25 angstrom structure of Xenopus Wnt8 (XWnt8) in complex with mouse Frizzled-8 (Fz8) cysteine-rich domain (CRD) reveals an unusual two-domain Wnt structure, not obviously related to known protein folds, resembling a “hand” with “thumb” and “index” fingers extended to grasp the Fz8-CRD at two distinct binding sites. One site is dominated by a palmitoleic acid lipid group projecting from serine 187 at the tip of Wnt’s thumb into a deep groove in the Fz8-CRD. In the second binding site, the conserved tip of Wnt’s “index finger” forms hydrophobic amino acid contacts with a depression on the opposite side of the Fz8-CRD. The conservation of amino acids in both interfaces appears to facilitate ligand-receptor cross-reactivity, which has important implications for understanding Wnt’s functional pleiotropy and for developing Wnt-based drugs for cancer and regenerative medicine.

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