Renalase's Expression and Distribution in Renal Tissue and Cells

Feng Wang,Zhonghua Zhao,Mei Bai,Niansong Wang,Zhigang Zhang,Tao Xing,Ruimin Hu,Shoufu Tian,Junhui Li,Utpal Sen

doi:10.1371/journal.pone.0046442

Abstract

To study renalase's expression and distribution in renal tissues and cells, renalase coded DNA vaccine was constructed, and anti-renalase monoclonal antibodies were produced using DNA immunization and hybridoma technique, followed by further investigation with immunological testing and western blotting to detect the expression and distribution of renalase among the renal tissue and cells. Anti-renalase monoclonal antibodies were successfully prepared by using DNA immunization technique. Further studies with anti-renalase monoclonal antibody showed that renalase expressed in glomeruli, tubule, mesangial cells, podocytes, renal tubule epithelial cells and its cells supernatant. Renalase is wildly expressed in kidney, including glomeruli, tubule, mesangial cells, podocytes and tubule epithelial cells, and may be secreted by tubule epithelial cells primarily.

Highlights

Renalase is a newly discovered monoamine oxidase enzyme originating from renal tissues [1]
We have been using recombinant renalase protein produced by prokaryotic expression system to develop monoclonal antibodies, and we have tried to use recombinant protein produced by eukaryotic expression systems such as baculovirus etc, to develop monoclonal antibodies
The results showed the recombinant renalase protein can be recognized by the monoclonal antibody (Fig. 2)

Summary

Introduction

Renalase is a newly discovered monoamine oxidase enzyme originating from renal tissues [1]. It degrades circulating catecholamines, regulates blood pressure and cardiac function, and is closely associated with cardiovascular diseases and chronic kidney disease (CKD) [2,3,4]. Renalase is a protein made of 342 amino acids with a molecular weight of 37.8 KDa approximately. Obtaining recombinant renalase protein and preparation of monoclonal antibodies are the essential steps for the study of renalase’s function, expression and distribution in renal tissues and cells. Obtaining large volume of eukaryotic expressed recombinant renalase protein has not been such an easy task [5,6]

Methods

Results

Conclusion