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Deletion mutants of the B type natriuretic peptide receptor: cDNA expression and protein purification and characterization

Abstract

The C type natriuretic peptide (CNP) is a peptide hormone stimulating vasorelaxation and inhibiting cell proliferation. CNP activates the type B natriuretic peptide receptor (NPR-B), known as the guanylate cyclase B membrane enzyme, which results in the cGMP release. To study functional properties of NPR-B, its gene fragments were expressed in methylotrophic yeastsPichia pastoris. Conditions were found providing for secretion of functionally active recombinant proteins NPR-Bs and NPR-Bl into the cultural medium in a yield of 25 mg/l culture. Their specific activity was 0.97 and 0.93 µmol cGMP min−1 mg−1 protein, respectively. It was shown that NPR-B belongs to the family of Ser/Thr protein kinases and can be autophosphorylated at the serine residues.

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Abbreviations

ANP:

BNP, and CNP, atrial, brain, and C type natriuretic peptides, respectively

AOX1:

alcohol oxidase 1

NPR-A:

natriuretic peptide receptor type A

NPR-B:

natriuretic peptide receptor type B

PMSF:

phenylmethylsufonyl chloride

YNB:

yeast nitrogenous base

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Correspondence to O. V. Solov’eva.

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Solov’eva, O.V., Kakuev, D.L., Gibanova, N.V. et al. Deletion mutants of the B type natriuretic peptide receptor: cDNA expression and protein purification and characterization. Russ J Bioorg Chem 26, 390–397 (2000). https://doi.org/10.1007/BF02758666

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  • DOI: https://doi.org/10.1007/BF02758666

Key words

  • bovine retina
  • heterologous expression
  • natriuretic peptide receptor
  • yeasts