Identification and characterization of collagen-like glycosylation and hydroxylation of CCN1.

Ishizawa, Yudai; Niwa, Yuki; Suzuki, Takehiro; Kawahara, Ryota; Dohmae, Naoshi; Simizu, Siro

Ishizawa, Yudai; Niwa, Yuki; Suzuki, Takehiro; Kawahara, Ryota; Dohmae, Naoshi; Simizu, Siro.

Afiliación

Ishizawa Y; Department of Applied Chemistry, Faculty of Science and Technology, Keio University, Yokohama 223-8522, Japan.
Niwa Y; Department of Applied Chemistry, Faculty of Science and Technology, Keio University, Yokohama 223-8522, Japan.
Suzuki T; Biomolecular Characterization Unit, RIKEN Center for Sustainable Resource Science, Wako 351-0198, Japan.
Kawahara R; Department of Applied Chemistry, Faculty of Science and Technology, Keio University, Yokohama 223-8522, Japan.
Dohmae N; Biomolecular Characterization Unit, RIKEN Center for Sustainable Resource Science, Wako 351-0198, Japan.
Simizu S; Department of Applied Chemistry, Faculty of Science and Technology, Keio University, Yokohama 223-8522, Japan.

Glycobiology ; 29(10): 696-704, 2019 09 20.

Article en En | MEDLINE | ID: mdl-31317175

RESUMEN

CCN1 is a secreted protein and belongs to the CCN family of matricellular proteins. CCN1 binds to various cell surface receptors; thus, CCN1 has important functions in cell proliferation, migration and angiogenesis through a variety of signaling pathways. We have reported that CCN1 is O-fucosylated and that this O-fucosylation regulates the secretion of CCN1 into the extracellular region. In this study, we detected collagen-like glycosylation and hydroxylation at Lys203 of recombinant CCN1 by mass spectrometry. We then examined the role of collagen-like glycosylation in the functions of CCN1. As a result, we found that a deficiency in collagen-like glycosylation decreased the secretion of CCN1 using wild-type CCN1- and collagen-like glycosylation-defective mutant CCN1-overexpressing cell lines. Further, knockout of lysyl hydroxylase3, a multifunctional protein with hydroxylase and glucosyltransferase activities, impaired the secretion and glycosylation level of recombinant CCN1. Previous studies reported that collagen glycosylation of Lys residues mediated by lysyl hydroxylase3 is glucosyl-galactosyl-hydroxylation, presuming that this collagen-like glycosylation detected at Lys203 of recombinant CCN1 in this study might be glucosyl-galactosyl-hydroxylation. Taken together, our results demonstrate the novel function of the collagen-like glycosylation of CCN1 and suggest that lysyl hydroxylase3-mediated glycosylation is important for CCN1 secretion.

Asunto(s)

Proteína 61 Rica en Cisteína/genética; Lisina/genética; Procolágeno-Lisina 2-Oxoglutarato 5-Dioxigenasa/genética; Receptores de Superficie Celular/genética; Secuencia de Aminoácidos/genética; Línea Celular; Movimiento Celular/genética; Proliferación Celular/genética; Colágeno/genética; Proteína 61 Rica en Cisteína/biosíntesis; Regulación de la Expresión Génica/genética; Glicosilación; Humanos; Hidroxilación; Espectrometría de Masas; Transducción de Señal/genética

Palabras clave

O-glycosylation; CCN1; hydroxylation; lysyl hydroxylase3; mass spectrometry

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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Procolágeno-Lisina 2-Oxoglutarato 5-Dioxigenasa / Receptores de Superficie Celular / Proteína 61 Rica en Cisteína / Lisina Tipo de estudio: Diagnostic_studies Límite: Humans Idioma: En Revista: Glycobiology Asunto de la revista: BIOQUIMICA Año: 2019 Tipo del documento: Article País de afiliación: Japón Pais de publicación: Reino Unido

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