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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
Authors: Hunt, R
Hettiarachchi, G
Katneni, U
Hernandez, N
Holcomb, D
Kames, J
Alnifaidy, R
Lin, B
Hamasaki-Katagiri, N
Wesley, A
Kafri, T
Morris, C
Bouche, L
Panico, M
Schiller, T
Ibla, J
Bar, H
Ismail, A
Morris, H
Komar, A
Kimchi-Sarfaty, C
Item Type: Journal Article
Abstract: 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.
Issue Date: 15-Nov-2019
Date of Acceptance: 13-Nov-2019
URI: http://hdl.handle.net/10044/1/75928
DOI: 10.3390/ijms20225734
ISSN: 1422-0067
Publisher: MDPI AG
Start Page: 1
End Page: 18
Journal / Book Title: International Journal of Molecular Sciences
Volume: 20
Issue: 22
Copyright Statement: © 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/).
Keywords: Science & Technology
Life Sciences & Biomedicine
Physical Sciences
Biochemistry & Molecular Biology
Chemistry, Multidisciplinary
Chemistry
ADAMTS13
synonymous variant
translation
codon usage
specific activity
post-translational modifications
ribosome profiling
CODON USAGE
O-FUCOSYLATION
PROTEIN
TRANSLATION
EXPRESSION
MUTATIONS
RNA
POLYMORPHISM
SUBSTRATE
ALTERS
ADAMTS13
codon usage
post-translational modifications
ribosome profiling
specific activity
synonymous variant
translation
Science & Technology
Life Sciences & Biomedicine
Physical Sciences
Biochemistry & Molecular Biology
Chemistry, Multidisciplinary
Chemistry
ADAMTS13
synonymous variant
translation
codon usage
specific activity
post-translational modifications
ribosome profiling
CODON USAGE
O-FUCOSYLATION
PROTEIN
TRANSLATION
EXPRESSION
MUTATIONS
RNA
POLYMORPHISM
SUBSTRATE
ALTERS
0399 Other Chemical Sciences
0604 Genetics
0699 Other Biological Sciences
Chemical Physics
Publication Status: Published
Article Number: ARTN 5734
Online Publication Date: 2019-11-15
Appears in Collections:Faculty of Natural Sciences