The majority of male infertility results from poor sperm motility. A direct link between altered protein phosphorylation and aberrant sperm motility has not been established. To address this issue, sperm samples obtained from 20 donors with healthy sperm and 20 donors with aberrantly motile sperm were subjected to computer assisted semen analysis (CASA), proteomic analysis, Western blot, and immunofluorescent staining. Proteomic analysis identified 12 protein spots as having differential phosphorylation, including gamma-tubulin complex associated protein 2 (GCP2). Western blot and immunofluorescence demonstrated differential expression of gamma-tubulin between healthy and aberrantly motile sperm. In conclusion, hypophosphorylated proteins and reduced expression of gamma-tubulin may be associated with low motility sperm.