Anthropogenic N-deposition represents a significant input of N into semi-arid chaparral and coastal sage scrub (CSS) shrublands of southern California. High levels of atmospheric N deposition have the potential to increase soil C and N mineralization, and we hypothesize that semi-arid shrubland soil exposed to long-term (decades) high N deposition will have significantly higher C and N mineralization potentials. This hypothesis was tested in a laboratory incubation where the inorganic N (NH4+NO3) and CO2 production of soils maintained at a constant temperature of 25°C and a soil moisture of 0.25 g H2O/g (65% water-filled pore space) were sampled sequentially over a 50-week period. The temporal trend in cumulative C and N mineralization was well described by a first- and zero-order model, respectively. Long-term atmospheric N deposition significantly increased potential N mineralization but not C mineralization, and both the rate and total N mineralization were significantly positively correlated with the surface (0–10 cm) soil δ 15N natural abundance and negatively correlated with the surface soil C:N ratio. While the incubation techniques used here do not provide realistic estimates of in situ C or N mineralization, these assays indicate that atmospheric N deposition has significantly altered ecosystem N storage and cycling.