Giant viruses infecting amoebae are notable for their substantial particle sizes and expansive genomes. In this study, we collected water samples from 44 distinct locations along a river within Brazil's Pantanal biome. These samples were processed and subsequently inoculated into Vermamoeba vermiformis cultures. Observation of cytopathic effects led to the isolation of a novel virus, designated Naiavirus, distinguished by its unprecedented virion morphology and genomic organization. To elucidate the protein composition of Naiavirus particles, large-scale infections were conducted using 20 T175 flasks, each containing 10 million amoebae. Post-infection, viral particles were purified through two successive sucrose gradient centrifugations to achieve high purity. Proteomic analysis of these purified particles identified 254 associated viral proteins, a count exceeding those reported for all known giant viruses except for pandoraviruses. This dataset encompasses the raw mass spectrometry data, identification files, and associated metadata, offering valuable insights into the structural proteome of this unique giant virus.