Cysteine sulfinate decarboxylase (CSD) which decarboxylates cysteine sulfinic acid (CSA) to form hypotaurine is thought to be involved in the biosynthesis of taurine. It was recently localized in astrocytes in the cerebellum and hippocampus by immunocytochemistry. Another sulfur-containing amino-acid (SCAA), homocysteic acid (HCA), was also found in astrocytes in these regions. We therefore investigated the specificity of CSD vs CSA and HCA as well as the related analogs homocysteine sulfinic acid (HCSA) and cysteic acid (CA). CSD was immunotrapped from brain and liver tissue supernatant using a specific CSD antiserum and Protein-A Sepharose. It was then incubated with the L-form of the various SCAA. Reaction products were identified and quantified by pre-column o-phthalaldehyde derivatization HPLC. CA and HCA from 2.5 to 25 mM inhibited the formation of hypotaurine from CSA (0.25 mM). Moreover, the inhibition curves were parallel for liver and brain CSD. CA or HCA (25 mM) elicited a near-total inhibition. HCSA did not produce a significant inhibition up to 25 mM. Incubation with 25 mM CSA or CA led to the formation of hypotaurine and taurine, respectively. The ratio of formation of taurine to that of hypotaurine was similar for CSD from liver and brain. In contrast no homotaurine, the decarboxylated reaction product of HCA, could be detected following incubation with 25 mM HCA. According to the sensitivity of the HPLC analysis this indicates that the decarboxylation of HCA, if any, was 130-fold and 50-fold less than that of CSA by CSD from liver and brain, respectively, in our experimental conditions. Similarly, following incubation with HCSA, no new peak appeared on the chromatogram when compared to a blank sample. These results show that CSD from either brain or liver has a high specificity for CSA and CA, which are the SCAA involved in the biosynthesis of taurine. HCA is an inhibitor of CSD but does not appear to be a substrate for CSD in vitro. HCSA is neither a substrate nor an inhibitor of CSD in vitro. Accordingly, CSD is unlikely to play a role in the metabolism of HCA or HCSA in vivo.