The aim of this study was to examine the differences between hydrophobicity and packing effects in specifying the three‐dimensional structure and stability of proteins when mutating hydrophobes in the hydrophobic core. In DNA‐binding proteins (leucine zippers), Leu residues are conserved at positions “d,” and β‐branched amino acids, Ile and Val, often occur at positions “a” in the hydrophobic core. In order to discern what effect this selective distribution of hydrophobes has on the formation and stability of two‐stranded α‐helical coiled coils/leucine zippers, three Val or three Ile residues were simultaneously substituted for Leu at either positions “a” (9, 16, and 23) or “d” (12, 19, and 26) in both chains of a model coiled coil. The stability of the resulting coiled coils was monitored by CD in the presence of Gdn·l. The results of the mutations of Ile to Val at either positions “a” or “d” in the reduced or oxidized coiled coils showed a significant hydrophobic effect with the additional methylene group in Ile stabilizing the coiled coil (δδG values range from 0.45 to 0.88 kcal/mol/mutation). The results of mutations of Leu to Ile or Val at positions “a” in the reduced or oxidized coiled coils showed a significant packing effect in stabilizing the coiled coil (δδG values range from 0.59 to 1.03 kcal/mol/mutation). Our results also indicate the subtle control hydrophobic packing can have not only on protein stability but on the conformation adopted by the amphipathic α‐helices. These structural findings correlate with the observation that in DNA‐binding proteins, the conserved Leu residues at positions “d” are generally less tolerant of amino acid substitutions than the hydrophobic residues at positions “a.”