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Molecular basis of sugar recognition by collectin-K1 and the effects of mutations associated with 3MC syndrome

BACKGROUND: Collectin-K1 (CL-K1, or CL-11) is a multifunctional Ca(2+)-dependent lectin with roles in innate immunity, apoptosis and embryogenesis. It binds to carbohydrates on pathogens to activate the lectin pathway of complement and together with its associated serine protease MASP-3 serves as a...

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Autores principales: Venkatraman Girija, Umakhanth, Furze, Christopher M, Gingras, Alexandre R, Yoshizaki, Takayuki, Ohtani, Katsuki, Marshall, Jamie E, Wallis, A Katrine, Schwaeble, Wilhelm J, El-Mezgueldi, Mohammed, Mitchell, Daniel A, Moody, Peter CE, Wakamiya, Nobutaka, Wallis, Russell
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4431178/
https://www.ncbi.nlm.nih.gov/pubmed/25912189
http://dx.doi.org/10.1186/s12915-015-0136-2
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author Venkatraman Girija, Umakhanth
Furze, Christopher M
Gingras, Alexandre R
Yoshizaki, Takayuki
Ohtani, Katsuki
Marshall, Jamie E
Wallis, A Katrine
Schwaeble, Wilhelm J
El-Mezgueldi, Mohammed
Mitchell, Daniel A
Moody, Peter CE
Wakamiya, Nobutaka
Wallis, Russell
author_facet Venkatraman Girija, Umakhanth
Furze, Christopher M
Gingras, Alexandre R
Yoshizaki, Takayuki
Ohtani, Katsuki
Marshall, Jamie E
Wallis, A Katrine
Schwaeble, Wilhelm J
El-Mezgueldi, Mohammed
Mitchell, Daniel A
Moody, Peter CE
Wakamiya, Nobutaka
Wallis, Russell
author_sort Venkatraman Girija, Umakhanth
collection PubMed
description BACKGROUND: Collectin-K1 (CL-K1, or CL-11) is a multifunctional Ca(2+)-dependent lectin with roles in innate immunity, apoptosis and embryogenesis. It binds to carbohydrates on pathogens to activate the lectin pathway of complement and together with its associated serine protease MASP-3 serves as a guidance cue for neural crest development. High serum levels are associated with disseminated intravascular coagulation, where spontaneous clotting can lead to multiple organ failure. Autosomal mutations in the CL-K1 or MASP-3 genes cause a developmental disorder called 3MC (Carnevale, Mingarelli, Malpuech and Michels) syndrome, characterised by facial, genital, renal and limb abnormalities. One of these mutations (Gly(204)Ser in the CL-K1 gene) is associated with undetectable levels of protein in the serum of affected individuals. RESULTS: In this study, we show that CL-K1 primarily targets a subset of high-mannose oligosaccharides present on both self- and non-self structures, and provide the structural basis for its ligand specificity. We also demonstrate that three disease-associated mutations prevent secretion of CL-K1 from mammalian cells, accounting for the protein deficiency observed in patients. Interestingly, none of the mutations prevent folding or oligomerization of recombinant fragments containing the mutations in vitro. Instead, they prevent Ca(2+) binding by the carbohydrate-recognition domains of CL-K1. We propose that failure to bind Ca(2+) during biosynthesis leads to structural defects that prevent secretion of CL-K1, thus providing a molecular explanation of the genetic disorder. CONCLUSIONS: We have established the sugar specificity of CL-K1 and demonstrated that it targets high-mannose oligosaccharides on self- and non-self structures via an extended binding site which recognises the terminal two mannose residues of the carbohydrate ligand. We have also shown that mutations associated with a rare developmental disorder called 3MC syndrome prevent the secretion of CL-K1, probably as a result of structural defects caused by disruption of Ca(2+) binding during biosynthesis. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12915-015-0136-2) contains supplementary material, which is available to authorized users.
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spelling pubmed-44311782015-05-15 Molecular basis of sugar recognition by collectin-K1 and the effects of mutations associated with 3MC syndrome Venkatraman Girija, Umakhanth Furze, Christopher M Gingras, Alexandre R Yoshizaki, Takayuki Ohtani, Katsuki Marshall, Jamie E Wallis, A Katrine Schwaeble, Wilhelm J El-Mezgueldi, Mohammed Mitchell, Daniel A Moody, Peter CE Wakamiya, Nobutaka Wallis, Russell BMC Biol Research Article BACKGROUND: Collectin-K1 (CL-K1, or CL-11) is a multifunctional Ca(2+)-dependent lectin with roles in innate immunity, apoptosis and embryogenesis. It binds to carbohydrates on pathogens to activate the lectin pathway of complement and together with its associated serine protease MASP-3 serves as a guidance cue for neural crest development. High serum levels are associated with disseminated intravascular coagulation, where spontaneous clotting can lead to multiple organ failure. Autosomal mutations in the CL-K1 or MASP-3 genes cause a developmental disorder called 3MC (Carnevale, Mingarelli, Malpuech and Michels) syndrome, characterised by facial, genital, renal and limb abnormalities. One of these mutations (Gly(204)Ser in the CL-K1 gene) is associated with undetectable levels of protein in the serum of affected individuals. RESULTS: In this study, we show that CL-K1 primarily targets a subset of high-mannose oligosaccharides present on both self- and non-self structures, and provide the structural basis for its ligand specificity. We also demonstrate that three disease-associated mutations prevent secretion of CL-K1 from mammalian cells, accounting for the protein deficiency observed in patients. Interestingly, none of the mutations prevent folding or oligomerization of recombinant fragments containing the mutations in vitro. Instead, they prevent Ca(2+) binding by the carbohydrate-recognition domains of CL-K1. We propose that failure to bind Ca(2+) during biosynthesis leads to structural defects that prevent secretion of CL-K1, thus providing a molecular explanation of the genetic disorder. CONCLUSIONS: We have established the sugar specificity of CL-K1 and demonstrated that it targets high-mannose oligosaccharides on self- and non-self structures via an extended binding site which recognises the terminal two mannose residues of the carbohydrate ligand. We have also shown that mutations associated with a rare developmental disorder called 3MC syndrome prevent the secretion of CL-K1, probably as a result of structural defects caused by disruption of Ca(2+) binding during biosynthesis. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12915-015-0136-2) contains supplementary material, which is available to authorized users. BioMed Central 2015-04-17 /pmc/articles/PMC4431178/ /pubmed/25912189 http://dx.doi.org/10.1186/s12915-015-0136-2 Text en © Venkatraman Girija et al.; licensee BioMed Central. 2015 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Venkatraman Girija, Umakhanth
Furze, Christopher M
Gingras, Alexandre R
Yoshizaki, Takayuki
Ohtani, Katsuki
Marshall, Jamie E
Wallis, A Katrine
Schwaeble, Wilhelm J
El-Mezgueldi, Mohammed
Mitchell, Daniel A
Moody, Peter CE
Wakamiya, Nobutaka
Wallis, Russell
Molecular basis of sugar recognition by collectin-K1 and the effects of mutations associated with 3MC syndrome
title Molecular basis of sugar recognition by collectin-K1 and the effects of mutations associated with 3MC syndrome
title_full Molecular basis of sugar recognition by collectin-K1 and the effects of mutations associated with 3MC syndrome
title_fullStr Molecular basis of sugar recognition by collectin-K1 and the effects of mutations associated with 3MC syndrome
title_full_unstemmed Molecular basis of sugar recognition by collectin-K1 and the effects of mutations associated with 3MC syndrome
title_short Molecular basis of sugar recognition by collectin-K1 and the effects of mutations associated with 3MC syndrome
title_sort molecular basis of sugar recognition by collectin-k1 and the effects of mutations associated with 3mc syndrome
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4431178/
https://www.ncbi.nlm.nih.gov/pubmed/25912189
http://dx.doi.org/10.1186/s12915-015-0136-2
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