Development of a High-Throughput Lysyl Hydroxylase (LH) Assay and Identification of Small-Molecule Inhibitors against LH2

Ashwini K. Devkota, John R. Veloria, Hou Fu Guo, Jonathan M. Kurie, Eun Jeong Cho, Kevin Dalby

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Lysyl hydroxylase-2 (LH2) catalyzes the hydroxylation of telopeptidyl lysine residues on collagen, leading to the formation of stable collagen cross-links that connect collagen molecules and stabilize the extracellular matrix. High levels of LH2 have been reported in the formation and stabilization of hydroxylysine aldehyde-derived collagen cross-links (HLCCs), leading to fibrosis and cancer metastasis in certain tissues. Identification of small-molecule inhibitors targeting LH2 activity requires a robust and suitable assay system, which is currently lacking. Thus, despite being a promising target for these diseases, small-molecule inhibitors for LH2 have yet to be reported. Therefore, we developed a luminescence-based strategy to monitor LH activity and validated its ability to identify new inhibitors in a screen of approximately 65,000 compounds against LH2. Primary hits were confirmed using the same LH assay against mimiviral L230. This newly developed LH assay is robust, suitable for high-throughput screening, and able to identify potent specific inhibitors of LH2.

Original languageEnglish (US)
Pages (from-to)484-491
Number of pages8
JournalSLAS Discovery
Volume24
Issue number4
DOIs
StatePublished - Apr 1 2019

Fingerprint

2-Oxoglutarate 5-Dioxygenase Procollagen-Lysine
Assays
Throughput
Molecules
Collagen
Hydroxylysine
Hydroxylation
Luminescence
Aldehydes
Lysine
Extracellular Matrix
Screening
Fibrosis
Stabilization

Keywords

  • cancer
  • high-throughput screen
  • luminescence
  • lysyl hydroxylase-2 (LH2)
  • succinate detection

ASJC Scopus subject areas

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Molecular Medicine

Cite this

Development of a High-Throughput Lysyl Hydroxylase (LH) Assay and Identification of Small-Molecule Inhibitors against LH2. / Devkota, Ashwini K.; Veloria, John R.; Guo, Hou Fu; Kurie, Jonathan M.; Cho, Eun Jeong; Dalby, Kevin.

In: SLAS Discovery, Vol. 24, No. 4, 01.04.2019, p. 484-491.

Research output: Contribution to journalArticle

Devkota, Ashwini K. ; Veloria, John R. ; Guo, Hou Fu ; Kurie, Jonathan M. ; Cho, Eun Jeong ; Dalby, Kevin. / Development of a High-Throughput Lysyl Hydroxylase (LH) Assay and Identification of Small-Molecule Inhibitors against LH2. In: SLAS Discovery. 2019 ; Vol. 24, No. 4. pp. 484-491.
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