The role of electrostatic interaction in the domain morphology of amide, ether, ester, and amine monoglycerol monolayers (abbreviated as ADD, ETD, ESD, and AMD, respectively) with systematic variation in the molecular structure of the headgroup region is investigated. Experimental studies using Brewster angle microscopy (BAM) and grazing incidence X-ray diffraction (GIXD) show that the characteristic features of the condensed monolayer phase, such as domain morphology, crystallinity, and lattice parameters, are very different for these monoglycerols. Therefore, the intermolecular interactions of the four amphiphilic monoglycerols are investigated in detail. First, the dipole moments of four monoglycerols of similar structure but with different functional groups are calculated by a semiempirical quantum mechanical technique. The dipole moments for monoglycerols follow the sequence AMD < ETD < ESD < ADD for the population of conformers of compounds investigated. The dipolar repulsion energies for the amphiphilic monoglycerols are also calculated for different possible mutual orientations between the dipoles. The calculated dipolar energies also follow the same trend for different possible headgroup orientations. These results can explain the domain shape of the monoglycerols observed experimentally. Second, ab initio calculations on the basis of the HF/6-31G** method are performed for representative monoglycerol headgroup segments. The results show that the intermolecular interaction energy related to dimer formation follows the order ETD < ESD < AMD < ADD segments, similar to that observed in experiment except in the case of the AMD segment. The relative importance of intra- and intermolecular hydrogen bonding in dimers is analyzed. The enhanced role of the intermolecular interaction relative to intramolecular interaction in the case of AMD contributes to the relatively high intermolecular interaction energy for the particular conformation of the dimer of AMD segment as observed from ab initio calculation. The present work shows that the variations in headgroup molecular structure alter drastically the domain shape, and the theoretical calculations conclusively reveal the important role of the electrostatic interactions for the mesoscopic domain architecture.