The kinetically unstable compound 3-mercapto-2-propenenitrile (HSCHCHCN) has been prepared for the first time by flash vacuum pyrolysis at 800 degrees C of 3-(tert-butylthio)-2-propenenitrile with a yield of 77% and a Z:E ratio of 8:1. Several deuterium and N-15 isotopologues were also prepared using isotopically enriched compounds. Quantum chemical calculations of the structural and conformational properties of the Z- and E-isomers were undertaken at the B3LYP/6-311++G(3df,2pd), MP2/6-311++G(3df,2pd), MP2/aug-cc-pVTZ, and G3 levels of theory. These methods all predict that the Z- and the E-forms each have two "stable" planar rotameric forms with the HSCC link of atoms in either a synperiplanar or an antiperiplanar conformation, with the synperiplanar form of the Z-isomer as the global minimum. The Z-isomer has been investigated by means of Stark-modulation microwave spectroscopy. Spectra attributable to the parent and three deuterium-substituted isotopologues of a single conformer were recorded and assigned. Additionally, the spectrum belonging to the first excited state of the lowest bending vibration was assigned. The ground-state rotational constants obtained by the least-squares analysis of these transitions were found to be in excellent agreement with the corresponding approximate equilibrium values generated by the MP2/aug-cc-pVTZ calculations. The preferred conformer of this molecule was found to have a synperiplanar arrangement of the HSCC chain of atoms and a planar or nearly planar geometry, with a stabilizing intramolecular hydrogen bond formed between the H atom of the thiol group and pi-electron density associated with the CN triple bond. The possible astrochemical/astrobiological significance of this compound is discussed.