Numerous reports have documented the interactions of fluorescent dissolved organic matter (FDOM) with other compounds such as metals and trace contaminants by characterizing the fluorescence quenching of the FDOM components. As FDOM is composed of numerous components, inter-component interactions can potentially take place. This study investigated the interactions between protein-like and humic-like components in FDOM using titration experiments and end-member mixing tests. We found that the co-occurrence of protein-like and humic-like components in FDOM samples resulted in an overlap behavior between their fluorescence peaks related to inter-component interactions. Our results suggest that the fluorescence of the protein-like components could be greatly quenched by the humic-like components in the FDOM samples, e.g., the humic-like components from Suwannee River and Nordic Reservoir FDOM yielded significant quenching effect for tyrosine (52% and 46%, respectively) and tryptophan (35% and 36%, respectively) in the titration experiments. The fluorescence of the humic-like components, however, was not impacted by the protein-like components. With the help of complexation modeling, we found that the binding capability between protein-like and humic-like components was dependent on their sources. This study could enhance our current knowledge on the role of FDOM in water and it is also important to the monitoring of FDOM by fluorescence spectroscopy.