Angiotensin II binding sites on micro-organisms contaminating cell cultures

An angiotensin II (Ang II) binding site, distinct from AT1 and AT2, has been found in cell cultures of rat aortic smooth muscle and rat glomerular mesangium. It is characterized by a high affinity for Ang II (Kd 0.75 +/- 0.13 nM) and Ang I (Ki 0.72 +/- 0.12 nM), but a very low affinity for Ang III (Ki 31 +/- 5 microM). Ang(1-7) (Ki 1.01 +/- 0.26 nM) and Ang(1-6) (Ki 4.54 +/- 0.24 nM) are very selective for this site, with affinities more than 150- and 10,000-fold greater, respectively, than for AT1 or AT2. The selective angiotensin receptor subtype ligands losartan and L-158,809 (AT1), PD 123319 and CGP 42112A (AT2) were inactive. Binding to this site was abolished after the cells had been treated with the antibiotic mixture BM-Cyclin, suggesting that the site is located not on the cells, but on a cell culture contaminant. This has been identified as Acholeplasma laidlawii. Caution should therefore be exercised when interpreting Ang II-related data obtained from cells that have not been checked for Mollicute contamination.