We have recently found that the computation time of homology-based subcellular localization can be substantially reduced by aligning profiles up to the cleavage site positions of signal peptides, mitochondrial targeting peptides, and chloroplast transit peptides [1]. While the method can reduce the profile alignment time by as much as 20 folds, it cannot reduce the computation time spent on creating the profiles. In this paper, we propose a new approach that can reduce both the profile creation time and profile alignment time. In the new approach, instead of cutting the profiles, we shorten the sequences by cutting them at the cleavage site locations. The shortened sequences are then presented to PSI-BLAST to compute the profiles. Experimental results and analysis of profile-alignment score matrices suggest that both profile creation time and profile alignment time can be reduced without sacrificing subcellular localization accuracy. Once a pairwise profile-alignment score matrix has b...