A scalable lysyl hydroxylase 2 expression system and luciferase-based enzymatic activity assay

Missense mutations that cause Bruck syndrome affect enzymatic activity, folding, and oligomerization of lysyl hydroxylase 2.

Bruck syndrome is a rare autosomal recessive connective tissue disorder characterized by fragile bones, joint contractures, scoliosis, and osteoporosis. The telopeptides of bone collagen I are underhydroxylated in these patients, leading to abnormal collagen cross-linking. Three point mutations in lysyl hydroxylase (LH) 2, the enzyme responsible for the hydroxylation of collagen telopeptides, h...

LightSwitchTM Luciferase Assay System

Lysyl hydroxylase

1 1 4 7 jci.org Volume 125 Number 3 March 2015 Introduction Collagen is the most abundant intrinsic matrix scaffolding protein and contributes to tissue tensile strength (1). Various aspects of collagen metabolism, including its expression, deposition, organization, and turnover, are aberrant in epithelial cancer and have been implicated in tumor progression (2). Evidence from multiple types of...

Suppression of fibroblast proliferation and lysyl hydroxylase activity by minoxidil.

The effect of minoxidil on lysyl hydroxylase activity and proliferation of human skin fibroblasts in culture was examined. Exposure of cells to minoxidil resulted in a specific loss of lysyl hydroxylase activity, the extent of which was dependent on the concentration of minoxidil from 25 to 500 microM and the duration of the treatment from 6 to 48 h. This phenomenon was unaffected by culture co...

Expression and Purification of the luciferase enzyme and in Vivo ATP Assay

Introduction: Gene expression and purification of luciferases from the firefly, Lampyris turkestanicus, and optimization of cellular ATP measurements were performed. Methods: cDNA encoding luciferases from Lampyris turkestanicus was transferred from pQE30 vector into pET28a expression vector and pLtu28 was built. Newly constructed vector was expressed in E. coli XL1 Blue and the recombinant l...