1. The metabolic cost and the mechanical work at different speeds during uphill, level and downhill walking have been measured in four subjects. 2. The mechanical work has been partitioned into the internal work (W(int)), due to the speed changes of body segment with respect to the body centre of mass (BCM), and the external work (W(ext)), related to the position and speed changes of the BCM in the environment. 3. W(ext) has been further divided into a positive part W+ext) and a negative one (W-(ext)), associated with the energy increases and decreases, respectively, over the stride period. 4. For all constant speeds the most economical gradient has been found to be -10.2% (+/- 0.8 S.D.). 5. At each gradient there is a unique W+ext/W-ext ratio (= 1 in level walking), regardless of speed, with a tendency for W-ext and W+ext to vanish above +15% and below -15% gradient, respectively. 6. W(int) is constant at each speed regardless of gradient. This is partly explained by an only slight decrease in stride frequency at increasing gradient. W(int) constancy implies that it has no role in determining the optimum gradient. 7. A linear multiple regression relating W+ext and W-ext to the metabolic cost at different gradients showed that negative (eff-) and positive (eff+) efficiencies decrease with increasing speed (from 0.912 to 0.726, and from 0.182 to 0.146, respectively). The eff-/eff+ ratio, however, remains rather constant (4.995 +/- 0.125 S.D.). 8. We conclude that the measured W(ext), the W+ext/W-ext partitioning and eff-/eff+ ratio, i.e. the different efficiency of the muscles used as force and brake generators, can explain the metabolic optimum gradient at about -10%.