The nature of the enhancing effect of muscle tissue on nonheme iron absorption in humans is unclear but thought to be related to muscle proteins. We conducted radioiron absorption studies to compare iron absorption from proteins isolated from beef and chicken muscle with that from freeze-dried beef and chicken muscle and from egg albumin. All meals contained an equivalent amount of protein as part of a semisynthetic liquid formula. Freeze-dried beef and chicken muscle increased iron absorption 180% (P < 0.001) and 100% (P < 0.001), respectively, relative to egg albumin. When added to the meal at an equivalent protein level (15 g), the isolated beef protein and the isolated heme-free beef protein with 94 and 98% protein content, respectively, increased iron absorption to the same extent as the native beef muscle. Similarly, when added to the meal at an equivalent protein level (30 g), isolated chicken muscle protein (94% protein) increased iron absorption similarly to native chicken muscle. Iron absorption from the meal containing the isolated heme-free chicken protein, however, was 120% (P < 0.01) greater than from the meal containing freeze-dried chicken muscle, indicating that a nonprotein component of muscle tissue with iron-binding potential may have been removed or concentrated by the protein extraction and separation procedures. Our results support the hypothesis that the enhancing effect of muscle tissue on iron absorption is mainly protein related but indicate that other factors may also play a role.