Legume crops are not usually fertilised with mineral N. However, there are at least two agronomic cases when it would be advantageous to distribute N fertiliser to legume crops: at sowing, before the onset of nodule functioning, and when a legume is intercropped with a cereal. We highlight the impact of various levels of fertiliser nitrogen on grain yield, nodulation capacity and biological nitrogen fixation in the four most common grain legume crops grown in central Italy. Chickpea (Cicer arietinum L.), field bean (Vicia faba L. var. minor), pea (Pisum sativum L.) and white lupin (Lupinus albus L.) were grown in soil inside growth boxes for two cropping seasons with five nitrogen fertilisation rates: 0, 40, 80, 120 and 160 kg ha−1. In both years, experimental treatments (five crops and five levels of N) were arranged in a randomised block design. We found that unfertilised plants overall yielded grain, total biomass and nitrogen at a similar level to plants supplied with 80–120 kg ha−1 of mineral nitrogen. However, above those N rates, the production of chickpea, pea and white lupin decreased, thus indicating that the high supply of N fertiliser decreased the level of N2 fixed to such an extent that the full N2-fixing potential might not be achieved. In all four grain legumes, the amount of N2 fixed was positively related to nodule biomass, which was inversely related to the rate of the N fertiliser applied. The four grain legumes studied responded differently to N fertilisation: in white lupin and chickpea, the amount of nitrogen derived from N2 fixation linearly decreased with increasing N supply as a result of a reduction in nodulation and N2 fixed per unit mass of nodules. Conversely, in field bean and pea, the decrease in N2 fixation was only due to a reduction in nodule biomass since nodule fixation activity increased with N supply. Our results suggest that the legume species and the N rate are critical factors in determining symbiotic N2-fixation responses to N fertilisation.