title: Presence of N-glycosylated transthyretin in plasma of V30M carriers in familial amyloidotic polyneuropathy: an escape from ERAD.
abstract: Familial amyloid polyneuropathy (FAP) is an autosomal dominant disease characterized by deposition of amyloid related to the presence of mutations in the transthyretin (TTR) gene. TTR is mainly synthesized in liver, choroid plexuses of brain and pancreas and secreted to plasma and cerebrospinal fluid (CSF). Although it possesses a sequon for N-glycosylation N-D-S at position 98, it is not secreted as a glycoprotein. The most common FAP-associated mutation is TTR V30M. In a screening for monoclonal antibodies developed against an amyloidogenic TTR form, we detected a distinct TTR with slower electrophoretic mobility in Western of plasma from carriers of the V30M mutation, not present in normal plasma. Mass spectrometry analyses of this slower migrating TTR (SMT) identified both wild-type and mutant V30M; SMT was undetectable upon N-glycosidase F treatment. Furthermore, SMT readily disappeared in the plasma of V30M - FAP patients after liver transplantation and appeared in plasma of transplanted domino individuals that received a V30M liver. SMT was also detected in plasma, but not in CSF of transgenic mice for the human V30M mutation. A hepatoma cell line transduced to express human V30M did not present the SMT modification in secretion media. Glycosylated TTR was absent in fibrils extracted from human kidney V30M autopsy tissue or in TTR aggregates extracted from the intestine of human TTR transgenic mice. Studies on the metabolism of this novel, glycosylated TTR secreted from FAP liver are warranted to provide new mechanisms in protein quality control and etiopathogenesis of the disease.
authors: Molecular Neurobiology, Instituto de Biologia Molecular e Celular, IBMC, Porto, Portugal.
authors: Anabela C
authors: AC
authors: Teixeira
authors: Maria J
authors: MJ
authors: Saraiva
mesh_terms: D028227:Amyloid Neuropathies, Familial / Q000097:blood* / Q000235:genetics* / Q000473:pathology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D060746:Endoplasmic Reticulum-Associated Degradation; D004797:Enzyme-Linked Immunosorbent Assay; D006031:Glycosylation; D006579:Heterozygote; D006801:Humans; D015151:Immunoblotting; D047468:Immunoprecipitation; D007413:Intestinal Mucosa / Q000378:metabolism; D007422:Intestines / Q000473:pathology; D007668:Kidney / Q000378:metabolism / Q000473:pathology; D008099:Liver / Q000378:metabolism / Q000473:pathology; D016031:Liver Transplantation; D013058:Mass Spectrometry; D051379:Mice; D008822:Mice, Transgenic; D009154:Mutation / Q000235:genetics*; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / Q000378:metabolism*; D011228:Prealbumin / Q000134:cerebrospinal fluid / Q000502:physiology*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D000911:Antibodies, Monoclonal; D011228:Prealbumin; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
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