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A Single Synonymous Variant (c.354G>A [p.P118P]) in ADAMTS13 Confers Enhanced Specific Activity
Synonymous variants within coding regions may influence protein expression and function. We have previously reported increased protein expression levels ex vivo (~120% in comparison to wild-type) from a synonymous polymorphism variant, c.354G>A [p.P118P], of the ADAMTS13 gene, encoding a plasma p...
Autores principales: | , , , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6888508/ https://www.ncbi.nlm.nih.gov/pubmed/31731663 http://dx.doi.org/10.3390/ijms20225734 |
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author | Hunt, Ryan Hettiarachchi, Gaya Katneni, Upendra Hernandez, Nancy Holcomb, David Kames, Jacob Alnifaidy, Redab Lin, Brian Hamasaki-Katagiri, Nobuko Wesley, Aaron Kafri, Tal Morris, Christina Bouché, Laura Panico, Maria Schiller, Tal Ibla, Juan Bar, Haim Ismail, Amra Morris, Howard Komar, Anton Kimchi-Sarfaty, Chava |
author_facet | Hunt, Ryan Hettiarachchi, Gaya Katneni, Upendra Hernandez, Nancy Holcomb, David Kames, Jacob Alnifaidy, Redab Lin, Brian Hamasaki-Katagiri, Nobuko Wesley, Aaron Kafri, Tal Morris, Christina Bouché, Laura Panico, Maria Schiller, Tal Ibla, Juan Bar, Haim Ismail, Amra Morris, Howard Komar, Anton Kimchi-Sarfaty, Chava |
author_sort | Hunt, Ryan |
collection | PubMed |
description | Synonymous variants within coding regions may influence protein expression and function. We have previously reported increased protein expression levels ex vivo (~120% in comparison to wild-type) from a synonymous polymorphism variant, c.354G>A [p.P118P], of the ADAMTS13 gene, encoding a plasma protease responsible for von Willebrand Factor (VWF) degradation. In the current study, we investigated the potential mechanism(s) behind the increased protein expression levels from this variant and its effect on ADAMTS13 physico-chemical properties. Cell-free assays showed enhanced translation of the c.354G>A variant and the analysis of codon usage characteristics suggested that introduction of the frequently used codon/codon pair(s) may have been potentially responsible for this effect. Limited proteolysis, however, showed no substantial influence of altered translation on protein conformation. Analysis of post-translational modifications also showed no notable differences but identified three previously unreported glycosylation markers. Despite these similarities, p.P118P variant unexpectedly showed higher specific activity. Structural analysis using modeled interactions indicated that subtle conformational changes arising from altered translation kinetics could affect interactions between an exosite of ADAMTS13 and VWF resulting in altered specific activity. This report highlights how a single synonymous nucleotide variation can impact cellular expression and specific activity in the absence of measurable impact on protein structure. |
format | Online Article Text |
id | pubmed-6888508 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-68885082019-12-09 A Single Synonymous Variant (c.354G>A [p.P118P]) in ADAMTS13 Confers Enhanced Specific Activity Hunt, Ryan Hettiarachchi, Gaya Katneni, Upendra Hernandez, Nancy Holcomb, David Kames, Jacob Alnifaidy, Redab Lin, Brian Hamasaki-Katagiri, Nobuko Wesley, Aaron Kafri, Tal Morris, Christina Bouché, Laura Panico, Maria Schiller, Tal Ibla, Juan Bar, Haim Ismail, Amra Morris, Howard Komar, Anton Kimchi-Sarfaty, Chava Int J Mol Sci Article Synonymous variants within coding regions may influence protein expression and function. We have previously reported increased protein expression levels ex vivo (~120% in comparison to wild-type) from a synonymous polymorphism variant, c.354G>A [p.P118P], of the ADAMTS13 gene, encoding a plasma protease responsible for von Willebrand Factor (VWF) degradation. In the current study, we investigated the potential mechanism(s) behind the increased protein expression levels from this variant and its effect on ADAMTS13 physico-chemical properties. Cell-free assays showed enhanced translation of the c.354G>A variant and the analysis of codon usage characteristics suggested that introduction of the frequently used codon/codon pair(s) may have been potentially responsible for this effect. Limited proteolysis, however, showed no substantial influence of altered translation on protein conformation. Analysis of post-translational modifications also showed no notable differences but identified three previously unreported glycosylation markers. Despite these similarities, p.P118P variant unexpectedly showed higher specific activity. Structural analysis using modeled interactions indicated that subtle conformational changes arising from altered translation kinetics could affect interactions between an exosite of ADAMTS13 and VWF resulting in altered specific activity. This report highlights how a single synonymous nucleotide variation can impact cellular expression and specific activity in the absence of measurable impact on protein structure. MDPI 2019-11-15 /pmc/articles/PMC6888508/ /pubmed/31731663 http://dx.doi.org/10.3390/ijms20225734 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Hunt, Ryan Hettiarachchi, Gaya Katneni, Upendra Hernandez, Nancy Holcomb, David Kames, Jacob Alnifaidy, Redab Lin, Brian Hamasaki-Katagiri, Nobuko Wesley, Aaron Kafri, Tal Morris, Christina Bouché, Laura Panico, Maria Schiller, Tal Ibla, Juan Bar, Haim Ismail, Amra Morris, Howard Komar, Anton Kimchi-Sarfaty, Chava A Single Synonymous Variant (c.354G>A [p.P118P]) in ADAMTS13 Confers Enhanced Specific Activity |
title | A Single Synonymous Variant (c.354G>A [p.P118P]) in ADAMTS13 Confers Enhanced Specific Activity |
title_full | A Single Synonymous Variant (c.354G>A [p.P118P]) in ADAMTS13 Confers Enhanced Specific Activity |
title_fullStr | A Single Synonymous Variant (c.354G>A [p.P118P]) in ADAMTS13 Confers Enhanced Specific Activity |
title_full_unstemmed | A Single Synonymous Variant (c.354G>A [p.P118P]) in ADAMTS13 Confers Enhanced Specific Activity |
title_short | A Single Synonymous Variant (c.354G>A [p.P118P]) in ADAMTS13 Confers Enhanced Specific Activity |
title_sort | single synonymous variant (c.354g>a [p.p118p]) in adamts13 confers enhanced specific activity |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6888508/ https://www.ncbi.nlm.nih.gov/pubmed/31731663 http://dx.doi.org/10.3390/ijms20225734 |
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