The assembly/disassembly process of carboxylated peptide-functionalized gold nanoparticles (peptide-GNPs) was studied within the pH interval of 2.5 to 10. The assembly process was not well controlled at pH 2.5, leading to the formation of 3D structures of GNPs, whereas at pH 4 we observed controlled assembly with the formation of only a network of 1D chains. In the pH range of 2.5 to 4, the assembly proceeded with the formation of a combination of two extremes (i.e., having both 1D and 2D nanostructures). The assembly process was reversed on changing the pH of the medium to 10. The assembly/disassembly process was monitored using UV-vis spectroscopy and finally confirmed by TEM analysis. This assembly resulted from the intermolecular H-bonding between two carboxylic acid groups of peptides bound to the two adjacent GNPs and were confirmed by FTIR spectroscopy.