The behavior of five polypeptides made of hydrophilic and pH-responsive aspartic acid (Asp) and hydrophobic phenylalanine (Phe), which had been prepared by stitching together short well-defined sequences of Asp and Phe, was studied as a function of pH. The effect of pH on these polypeptides referred to as (Asp(3)Phe(1))(n), (Asp(2)Phe(1))(n), (Asp(1)-Phe(1))(n), (Asp(1),Phe(2))(n), and (Asp(1)Phe(3))(n), varied dramatically depending on their constituting sequence. The more hydrophobic polypeptides (Asp(1)Phe(2))(n) and (Asp(1)Phe(3))(n) behaved as if the Asp's were isolated from each other and showed an apparent PKa (pK(a)(aPP)) that remained constant with level of ionization (alpha = [Asp(-1)]/[Asp](total)) and equaled 5.4 and 6.4, respectively. The more hydrophilic polypeptides (Asp(3)Phe(1))(n) and (Asp(2)Phe(1))(n) behaved like weak polyacids showing a linear increase in pKaaPP with increasing a. The pKaaPP of (Asp(1)Phe(1))(n) showed a trend as a function of alpha intermediate between the Asp-rich and Phe-rich polypeptides, behaving as if the Asp's were isolated at low a values (<0.35) but acting as a weak polyacid for large a values (>0.35). The effect that a, and thus the charge density of the polypeptides, had on the collapse and aggregation of the polypeptides was characterized by conducting static light scattering and fluorescence measurements. Static light scattering measurements demonstrated that all polypeptides precipitated and aggregated in solution at a critical charge density of 0.2. Fluorescence measurements with pyrene indicated that this behavior was due to the formation of Phe aggregates in water. Together, these experiments provide a complete description of how pH affects the behavior of a series of unique amphiphilic polypeptides designed with a well-defined sequence.