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Hedgehog proteins create a dynamic cholesterol interface

Mafi, Amirhossein and Purohit, Rahul and Vielmas, Erika and Lauinger, Alexa R. and Lam, Brandon and Cheng, Yu-Shiuan and Zhang, Tianyi and Huang, Yiran and Kim, Soo-Kyung and Goddard, William A., III and Ondrus, Alison E. (2021) Hedgehog proteins create a dynamic cholesterol interface. PLoS ONE, 16 (2). Art. No. e0246814. ISSN 1932-6203. https://resolver.caltech.edu/CaltechAUTHORS:20210301-120802550

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Abstract

During formation of the Hedgehog (Hh) signaling proteins, cooperative activities of the Hedgehog INTein (Hint) fold and Sterol Recognition Region (SRR) couple autoproteolysis to cholesterol ligation. The cholesteroylated Hh morphogens play essential roles in embryogenesis, tissue regeneration, and tumorigenesis. Despite the centrality of cholesterol in Hh function, the full structure of the Hint-SRR (“Hog”) domain that attaches cholesterol to the last residue of the active Hh morphogen remains enigmatic. In this work, we combine molecular dynamics simulations, photoaffinity crosslinking, and mutagenesis assays to model cholesterolysis intermediates in the human Sonic Hedgehog (hSHH) protein. Our results provide evidence for a hydrophobic Hint-SRR interface that forms a dynamic, non-covalent cholesterol-Hog complex. Using these models, we suggest a unified mechanism by which Hh proteins can recruit, sequester, and orient cholesterol, and offer a molecular basis for the effects of disease-causing hSHH mutations.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1371/journal.pone.0246814DOIArticle
ORCID:
AuthorORCID
Mafi, Amirhossein0000-0002-8366-6785
Purohit, Rahul0000-0001-9712-4277
Vielmas, Erika0000-0003-0263-7278
Lauinger, Alexa R.0000-0002-9820-9130
Huang, Yiran0000-0002-1435-6281
Kim, Soo-Kyung0000-0002-4498-5441
Goddard, William A., III0000-0003-0097-5716
Ondrus, Alison E.0000-0002-6023-3290
Additional Information:© 2021 Mafi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Received: October 22, 2020; Accepted: January 26, 2021; Published: February 25, 2021. We thank B Lomenick and the Caltech Proteome Exploration Laboratory for help with mass spectrometry experiments, DS-W Peng for assistance with hSHH mutant analysis, and GW Mobbs and A Hoelz for plasmids, reagents, and advice on in vitro protein expression. Data Availability Statement: All relevant data are within the manuscript and its Supporting Information files. Funding: The Margaret E. Early Medical Research Trust provided funding for AM, S-KK, and WAG. The authors have declared that no competing interests exist. Author Contributions: Conceptualization: Amirhossein Mafi, Rahul Purohit, Soo-Kyung Kim, William A. Goddard, III, Alison E. Ondrus. Data curation: Amirhossein Mafi, Yiran Huang, Soo-Kyung Kim, Alison E. Ondrus. Formal analysis: Amirhossein Mafi, Rahul Purohit, Brandon Lam, Yu-Shiuan Cheng, Yiran Huang, Alison E. Ondrus. Investigation: Amirhossein Mafi, Rahul Purohit, Erika Vielmas, Alexa R. Lauinger, Yu-Shiuan Cheng, Tianyi Zhang, Yiran Huang, Soo-Kyung Kim, William A. Goddard, III, Alison E. Ondrus. Methodology: Amirhossein Mafi, Rahul Purohit, Erika Vielmas, Tianyi Zhang, William A. Goddard, III, Alison E. Ondrus. Project administration: Rahul Purohit, Erika Vielmas, Yu-Shiuan Cheng, Soo-Kyung Kim, William A. Goddard, III, Alison E. Ondrus. Resources: Erika Vielmas, William A. Goddard, III. Software: William A. Goddard, III. Supervision: Rahul Purohit, Erika Vielmas, Soo-Kyung Kim, William A. Goddard, III, Alison E. Ondrus. Validation: Amirhossein Mafi, Rahul Purohit, Erika Vielmas, Alexa R. Lauinger, Brandon Lam. Visualization: Amirhossein Mafi, Alison E. Ondrus. Writing – original draft: Amirhossein Mafi, Alison E. Ondrus. Writing – review & editing: Rahul Purohit, Erika Vielmas, Alexa R. Lauinger, Yu-Shiuan Cheng, Tianyi Zhang, William A. Goddard, III.
Funders:
Funding AgencyGrant Number
Margaret E. Early Medical Research TrustUNSPECIFIED
Issue or Number:2
Record Number:CaltechAUTHORS:20210301-120802550
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20210301-120802550
Official Citation:Mafi A, Purohit R, Vielmas E, Lauinger AR, Lam B, Cheng Y-S, et al. (2021) Hedgehog proteins create a dynamic cholesterol interface. PLoS ONE 16(2): e0246814. https://doi.org/10.1371/journal.pone.0246814
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:108248
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:01 Mar 2021 20:29
Last Modified:01 Mar 2021 20:29

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