ADAMTS7: Recombinant Protein Expression and Purification

Methods Mol Biol. 2020:2043:63-73. doi: 10.1007/978-1-4939-9698-8_6.

Abstract

ADAMTS7 is a secreted protease that is predominantly expressed in tissues of the cardiovascular system and tendon. Although recent evidence suggests that it may play a role in the etiology of coronary artery disease, its physiological function and substrates are unknown. The enzyme undergoes extensive posttranslational modifications, including chondroitin sulfate attachment, N and O-linked glycosylation, and a two-step activation process. For the benefit of scientists who study the function of ADAMTS7 and its role in disease, this chapter provides an introduction to the chemical and functional properties of the various ADAMTS7 domains, as well as a protocol for the recombinant expression and purification of ADAMTS7.

Keywords: ADAMTS7; Chondroitin sulfate; Ion exchange chromatography; Posttranslational modification; Protein purification; Proteoglycan; Recombinant protein expression; Transient transfection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ADAMTS7 Protein / chemistry
  • ADAMTS7 Protein / genetics
  • ADAMTS7 Protein / metabolism
  • Crystallography, X-Ray
  • Glycosylation
  • HEK293 Cells
  • Humans
  • Models, Molecular
  • Protein Domains
  • Protein Engineering / methods*
  • Protein Processing, Post-Translational
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism

Substances

  • Recombinant Proteins
  • ADAMTS7 Protein
  • ADAMTS7 protein, human