Glycosylation is critical for natriuretic peptide receptor-B function

Co-transfection of a truncated natriuretic peptide receptor-B (NPR-B) with the full length receptor results in a decrease of 60–80% in wild-type receptor activity. This reduction correlates with a loss of glycosylation of the full length NPR-B. This effect is dose-dependent, and occurs with no change in the glycosylation of the truncated receptor. Co-transfection of the full length NPR-B with other receptors yields similar results. These data suggest that glycosylation may be crucial for NPR-B function. Cross-linking studies further demonstrate that only fully glycosylated NPR-B receptors are able to bind ligand. Our data therefore argue that carbohydrate modification may be critical for NPR-B receptor ligand binding.Abbreviations as amino acids - ANF atrial natriuretic factor - ANOVA analysis of variance - BS³ bis(sulfosuccinimidyl) suberate - BSA bovine serum albumin - CNP C-type natriuretic peptide - DEAE dextran-diethylaminoethyl-dextran - DMEM Dulbecco's modified Eagle medium - DMSO dimethyl sulfoxide - dNTP deoxynucleotide triphosphate - EDTA ethylenediamine tetraacetic acid - IBMX 3-isobutyl-l-methyl-=xanthine - min minutes - N-linked asparagine-linked - NPR natriuretic peptide receptor - nt nucleotide - PCR polymerase chain reaction - RIA radioimmunoassay - RP-HPLC reverse phase-high performance liquid chromatography - RP-HPLC reverse phase-high performance liquid chromatography - SDS sodium dodecyl sulfate - UV ultraviolet Address for offprints:Department of Pharmacology, University of Montreal, 2900 Edouard Montpetit, Montreal, Quebec, H3C3J7, Canada