title: Novel mutations in exon 2 of MATN3 affect residues within the alpha-helices of the A-domain and can result in the intracellular retention of mutant matrilin-3.
abstract: Multiple epiphyseal dysplasia (MED) is a clinically variable and genetically heterogeneous chondrodysplasia characterized by mild to moderate short stature and early onset osteoarthritis. Some forms of MED result from mutations in the gene encoding the cartilage structural protein matrilin-3 (MATN3). The majority of MATN3 mutations affect conserved residues within the beta-sheet of the single A-domain of matrilin-3. These mutations cause the protein to misfold and prevent its secretion from the rER, both in vitro and in vivo. More recently a single mutation (p.Phe105Ser) has been identified within the alpha1-helix of the A-domain, but its affect on the structure and/or function of matrilin-3 is unknown. In this paper we describe the characterization of two additional alpha-helical mutations (p.Ala173Asp and p.Lys231Asn) and show that both p.Phe105Ser and pAla173Asp prevent the secretion of A-domain in vitro. In contrast, p.Lys231Asn does not prevent the secretion of matrilin-3 A-domain, nor does it disrupt the structure of this domain or inhibit its binding to type II or type IX collagen. Therefore, despite extensive biochemical analysis the disease mechanism of p.Lys231Asn remains unresolved and care should be taken in counseling for these types of mutation in MATN3.
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester, United Kingdom.
authors.forename: Maryline
authors.initials: M
authors.lastname: Fresquet
authors.forename: Gail C
authors.initials: GC
authors.lastname: Jackson
authors.forename: John
authors.initials: J
authors.lastname: Loughlin
authors.forename: Michael D
authors.initials: MD
authors.lastname: Briggs
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D002675:Child, Preschool; D024043:Collagen Type II / Q000378:metabolism; D024081:Collagen Type IX / Q000378:metabolism; D004252:DNA Mutational Analysis; D005091:Exons / Q000235:genetics*; D016326:Extracellular Matrix Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D064235:Matrilin Proteins; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000378:metabolism*; D009154:Mutation / Q000235:genetics*; D010009:Osteochondrodysplasias / Q000235:genetics; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D011485:Protein Binding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D024043:Collagen Type II; D024081:Collagen Type IX; D016326:Extracellular Matrix Proteins; D064235:Matrilin Proteins; D050505:Mutant Proteins; D011994:Recombinant Proteins