A Single Synonymous Variant (c.354G&gt;A [p.P118P]) in ADAMTS13 Confers Enhanced Specific Activity

Hemostasis Branch, Division of Plasma Protein Therapeutics, Office of Tissues and Advanced Therapies, Center for Biologics Evaluation & Research, US FDA, Silver Spring, MD 20993, USA

Gene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA

Present Address: Department of Emergency Medicine, Banner University Medical Center, The University of Arizona, Tucson, AZ 85724, USA

⁴

BioPharmaSpec Ltd., St. Saviour JE2 7LA, UK

⁵

Department of Life Sciences, Imperial College London, South Kensington Campus, London SW7 2AZ, UK

⁶

Present Address: Antikor Biopharma Ltd., Stevenage Bioscience Catalyst, Gunnels Wood Road, Stevenage SG1 2FX, UK

⁷

Departments of Cardiac Surgery and Anesthesiology, Perioperative and Pain Medicine, Boston Children’s Hospital and Harvard Medical School, Boston, MA 02115, USA

⁸

Department of Statistics, University of Connecticut, Storrs, CT 06269, USA

⁹

Center for Gene Regulation in Health and Disease, Department of Biological, Geological & Environmental Sciences, Cleveland State University, Cleveland, OH 44115, USA

Author to whom correspondence should be addressed.

^†

These authors contributed equally to this work.

Int. J. Mol. Sci. 2019, 20(22), 5734; https://doi.org/10.3390/ijms20225734

Received: 8 October 2019 / Revised: 9 November 2019 / Accepted: 13 November 2019 / Published: 15 November 2019

(This article belongs to the Collection Feature Papers in Molecular Biology)

View Full-Text Download PDF

Browse Figures

Citation Export

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. View Full-Text

Keywords: ADAMTS13; synonymous variant; translation; codon usage; specific activity; post-translational modifications; ribosome profiling

▼ Show Figures

Figure 1

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited

Supplementary Material

Supplementary File 1:
PDF-Document (PDF, 1130 KiB)

Share and Cite

MDPI and ACS Style

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.; Bouché, L.; Panico, M.; Schiller, T.; Ibla, J.; Bar, H.; Ismail, A.; Morris, H.; Komar, A.; Kimchi-Sarfaty, C. A Single Synonymous Variant (c.354G>A [p.P118P]) in ADAMTS13 Confers Enhanced Specific Activity. Int. J. Mol. Sci. 2019, 20, 5734. https://doi.org/10.3390/ijms20225734

AMA Style

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, Bouché L, Panico M, Schiller T, Ibla J, Bar H, Ismail A, Morris H, Komar A, Kimchi-Sarfaty C. A Single Synonymous Variant (c.354G>A [p.P118P]) in ADAMTS13 Confers Enhanced Specific Activity. International Journal of Molecular Sciences. 2019; 20(22):5734. https://doi.org/10.3390/ijms20225734

Chicago/Turabian Style

Hunt, Ryan, Gaya Hettiarachchi, Upendra Katneni, Nancy Hernandez, David Holcomb, Jacob Kames, Redab Alnifaidy, Brian Lin, Nobuko Hamasaki-Katagiri, Aaron Wesley, Tal Kafri, Christina Morris, Laura Bouché, Maria Panico, Tal Schiller, Juan Ibla, Haim Bar, Amra Ismail, Howard Morris, Anton Komar, and Chava Kimchi-Sarfaty. 2019. "A Single Synonymous Variant (c.354G>A [p.P118P]) in ADAMTS13 Confers Enhanced Specific Activity" International Journal of Molecular Sciences 20, no. 22: 5734. https://doi.org/10.3390/ijms20225734

Find Other Styles

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Metrics

Abstract views Pdf views Html views

Article Menu

A Single Synonymous Variant (c.354G>A [p.P118P]) in ADAMTS13 Confers Enhanced Specific Activity

Abstract

Supplementary Material

Share and Cite

Article Metrics

Article Access Statistics

Article Access Map by Country/Region

Further Information

Guidelines

MDPI Initiatives

Follow MDPI