Renalase is a recently discovered protein, involved in regulation of blood pressure in humans and animals. Although several splice variants of human renalase mRNA transcripts have been recognized, only one protein product, hRenalase1, has been found so far. In this study, we have used polymerase chain reaction (PCR)-based amplification of individual exons of the renalase gene and their joining for construction of full-length hRenalase2 coding sequence followed by expression of hRenalase2 as a polyHis recombinant protein in Escherichia coli cells. To date this is the first report on synthesis and purification of hRenalase2. Applicability of this approach was verified by constructing hRenalase1 coding sequence, its sequencing and expression in E. coli cells. hRenalase1 was used for generation of polyclonal antiserum in sheep. Western blot analysis has shown that polyclonal anti-renalase1 antibodies effectively interact with the hRenalase2 protein. The latter suggests that some functions and expression patterns of hRenalase1 documented by antibody-based data may be attributed to the presence of hRenalase2. The realized approach may be also used for construction of coding sequences of various (especially weakly expressible) genes, their transcript variants, etc.
BackgroundRenalase is a recently discovered secretory protein involved in regulation of arterial blood pressure in humans and animals. Results of animal experiments from independent laboratories indicate that administration of human recombinant renalase decreases blood pressure and some genetically predisposed hypertensive rats have lowered renalase levels.Material/MethodsThe levels of renalase mRNA expression in brain hemispheres, heart, and kidneys of spontaneously hypertensive rats (SHR) with moderate (140–180 mm Hg) or high (>180 mm Hg) hypertension and of control Wistar-Kyoto (WKY) rats were analyzed using real-time PCR.ResultsSpontaneously hypertensive rats with high hypertension (>180 mm Hg) had a lower renalase mRNA level in brain hemispheres, and higher heart and kidney renalase mRNA levels compared with control WKY rats. In SHR with a moderate increase in arterial blood pressure (140–180 mm Hg), the tissue renalase mRNA changed in the same direction but did not reach the level of statistical significance as compared with control rats.ConclusionsThe results indicate that the development of hypertension in SHR is accompanied by altered expression of the renalase gene in the examined organs as compared with control WKY rats. The brain and peripheral tissues renalase mRNA levels demonstrate opposite trends, which are obviously crucial for impaired regulation of blood pressure in SHR.
Renalase is a recently discovered secretory enzyme responsible for selective degradation of blood catecholamines. The review summarizes literature data on expression of this enzyme and on its structure and functions. Special attention is paid to unsolved and questionable problems including: 1) prediction of the presence of FAD in the protein structure based on amino acid sequence similarity of renalase with known FAD-dependent enzymes; 2) identity of plasma and urinary renalase; 3) mechanism underlying conversion of inactive renalase into the active form.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.