Lipid Transfer Protein I is recognized as a key component involved in high density lipoprotein metabolism. We have been studying lipid transfer in relation to sperm capacitation, a complex series of cell surface events required for the acrosome reaction and fertilization. We have previously shown that Lipid Transfer Protein I is present and active in the female reproductive tract. In the present study, we show that purified Lipid Transfer Protein I directly stimulates human sperm capacitation, but not the acrosome reaction, in the absence of other biological effectors. These results provide strong evidence for a novel role for Lipid Transfer Protein I and reveal, for the first time, a potent activator of capacitation, prior to the acrosome reaction.