Expression of catalytically active recombinant lysyl oxidase and lysyl oxidase-like 2 by mammalian cells in cell medium

Date
2014
DOI
Authors
Bhatnagar, Vani
Version
OA Version
Citation
Abstract
The lysyl oxidase (LOX) family of proteins consist of copper-dependent amine oxidases that play critical roles in the biogenesis of connective tissue matrices by crosslinking the extracellular matrix proteins, collagen and elastin. The present study expresses recombinant LOX and LOX-like2 (LOXL2) in mammalian cells (MDA-MB23 l and CHO-K 1 respectively) as catalytically active enzymes in the medium. After normalizing to the protein concentration present in medium, it was found that the enzyme acitivity of the recombinant LOXL2 expressed by the transduced CHO-Kl cells was 1.4 fold that of the non-transduced control cells and the enzyme activity of the recombinant LOX expressed by the transduced MDA-MB231 cells was 15.9 times that of the non-transduced control cells. Enzymatic activity was supported by the expression of mature LOX at 37 kDa in the medium of the transduced MDA-MB23 l cells and the expression ofLOXL2 at 87 kDa in the medium of the transduced CHO-Kl cells. Availability of high yield pure and active forms of recombinant LOX and LOXL2 would be significantly helpful in functional studies related to substrate specificity and crystal structures of this family of amine oxidases.
Description
PLEASE NOTE: This work is protected by copyright. Downloading is restricted to the BU community: please click Download and log in with a valid BU account to access. If you are the author of this work and would like to make it publicly available, please contact open-help@bu.edu.
Thesis (MSD) --Boston University, Henry M. Goldman School of Dental Medicine, 2014 (Department of Oral Biology).
Includes bibliography: leaves 50-51.
License
This work is protected by copyright. Downloading is restricted to the BU community. If you are the author of this work and would like to make it publicly available, please contact open-help@bu.edu.