Effect of Specific Mutations in Cd300 Complexes Formation; Potential Implication of Cd300f in Multiple Sclerosis
Martínez-Barriocanal, Águeda (Centro de Investigación Biomédica en Red de Bioingeniería, Biomateriales y Nanomedicina)
Arcas-García, Andrea (Hospital Universitari Vall d'Hebron. Institut de Recerca)
Magallon-Lorenz, Miriam (Hospital Universitari Vall d'Hebron. Institut de Recerca)
Ejarque-Ortiz, Aroa (Hospital Universitari Vall d'Hebron. Institut de Recerca)
Negro-Demontel, María Luciana (Neuroinflammation and Gene Therapy Laboratory, Institut Pasteur Montevideo)
Comas-Casellas, Emma (Hospital Universitari Vall d'Hebron. Institut de Recerca)
Schwartz, Simo (Centro de Investigación Biomédica en Red de Bioingeniería, Biomateriales y Nanomedicina)
Malhotra, Sunny (Hospital Universitari Vall d'Hebron)
Montalban, Xavier (Hospital Universitari Vall d'Hebron. Institut de Recerca)
Peluffo, Hugo (UDELAR)
Martín, Margarita (Institut d'Investigacions Biomèdiques August Pi i Sunyer)
Comabella, Manuel
Sayós, Joan (Centro de Investigación Biomédica en Red de Bioingeniería, Biomateriales y Nanomedicina)
Universitat Autònoma de Barcelona

Data:	2017
Resum:	Herein, we have used bioinformatics tools to predict five clusters defining ligand-binding sites on the extracellular domain of human CD300b receptor, presumably involved in the formation of both homodimers and heterodimers with other CD300 family members. Site-directed mutagenesis revealed residues glutamic acid 28 and glutamine 29 in cluster 5 to be necessary for the formation of CD300b complexes. Surprisingly, the disruption of cluster 2 and 4 reconstituted the binding capability lost by the mutation of residues glutamic acid 28 to alanine, glutamine 29 to alanine (E28A-Q29G). We identified a missense mutation arginine 33 to glutamine (R33Q) in CD300f by direct sequencing of exon 2 in peripheral blood samples from 50 patients with multiple sclerosis (MS). Levels of expression of CD300f were almost undetectable on monocytes from the patient bearing the R33Q mutation compared with healthy individuals. Whereas R33Q mutation had no effect in the formation of CD300f complexes, the inhibition of protein synthesis with cycloheximide indicated that CD300f R33Q is less stable than native CD300f. Finally, we report that the levels of expression of CD300f on the surface of classical and intermediate monocytes from MS patients are significantly lower when compared to the same cell populations in healthy individuals.
Ajuts:	Ministerio de Ciencia e Innovación PI1100045
Drets:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Llengua:	Anglès
Document:	Article ; recerca ; Versió publicada
Publicat a:	Scientific reports, Vol. 7 (october 2017) , ISSN 2045-2322

DOI: 10.1038/s41598-017-12881-8
PMID: 29051512

11 p, 3.7 MB

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