Immunoactive iso-ANP/BNP in plasma, tissues and atrial granules of the rat

We have developed a specific radioimmunoassay (RIA) for iso-rANP(1-45)/rBNP(51-95) using antiserum produced against a peptide comprise of the first 20 amino acid sequence of this cardiac hormone. Using this RIA, we found that atria contained the highest amount of iso-rANP(1-45) (567.37 pmol/g) which is about 140-fold higher than ventricles (4.32 pmol/g). With the exception of the lung and kidney, all other tissues had negligible amounts. The plasma level was 1.4 fmol/ml and the only molecular form found was the 45 amino acid form. HPLC analysis of extracts of isolated, purified atrial granules revealed that, like atrial natriuretic peptide (ANP), iso-rANP/rBNP is also stored in these granules. However, while ANP is stored predominantly as pro-ANP(1-126) and cleaved during or after release, iso-ANP/BNP is stored as the 45 amino acid form and may be processed prior to storage in the granules.