Metabolic depletion of hereditary ovalocytes leads, similar to normal red cells, to decreased intracellular concentrations of ATP and GSH as well as degradation of the phosphoinositides to phosphatidylinositol and diacylglycerol. In contrast to normal red cells, however, loss of ATP does not induce any gross shape transformations; even after extensive depletion the ovalocytes retain their initial elongated stomatocytic character. The mechanical properties of hereditary ovalocytes are associated with a deletion of nine amino acid residues in band 3. Since the deletion appears to increase the stiffness of a normally flexible region of band 3, connecting the N-terminal cytoplasmic domain with the membrane spanning domain, our results indicate that shape changes require a flexible attachment of the cytoskeleton to the membrane-spanning band 3. The results also imply that metabolism of phosphoinositide cannot be the only determinant of cell shape, as suggested by the bilayer-couple hypothesis, but also other factors are involved in metabolically induced shape transformations.