Midbody, a transient organelle-like structure, is known as central for abscission and is indispensable for termination of cytokinesis. Here, we used the midbody proteome inventories to construct the potential midbody protein-protein interaction (PPI) network. To delineate novel regulators participating in cytokinesis, the z-score, a standard statistic score, rather than hub degree was implemented to prioritize the novel hubs. Of these hubs, KIAA0133, SEPT1, KIAA1377, and CRMP-1 were localized to the midbody, whereas HTR3A and ICAM2 were associated with the cleavage furrow as examined by immunofluorescence. Knockdown of SEPT1 and KIAA1377 resulted in increasing numbers of cytokinesis defect cells, suggesting these newly identified hubs play critical roles in cytokinesis progression. Moreover, ectopic expression of CRMP-1 mutant in which Aurora-A phosphorylation sites have been replaced with Ala results in a cytokinesis defect. This subproteome network construction not only sheds light on the intimate interactions of the midbody proteomes, but also prioritizes novel hubs or protein complexes that may govern the process of cytokinesis.