Evolutionary and Structural Assessment of the Human Secreted Frizzled-Related Protein (SFRP) Family

It has been observed that five members of Secreted Frizzled-Related proteins act as antagonists for the Wnt signaling pathway in humans. These glycoproteins have two functional domains: the cysteine-rich domain (CRD) and the netrin-related domain (NTR), with a completely conserved disulfide bond in the CRD domain. Phylogenetic analysis revealed that this protein family can be divided into two subgroups, SFRP1/SFRP2/SFRP5 versus SFRP3/SFRP4. The SFRP3/SFRP4 group was found to be more closely related to the sponge Lubomirskia baicalensis, which is believed to represent the ancient origin of SFRPs. The model evaluation demonstrated high-quality conformational homology modeling in the predicted Human SFRP models compared to the Sizzled crystal structure of Xenopus laevis. The molecular dynamic simulation illustrated that SFRP1 and SFRP2 exhibit the most stable structures during 100 ns of simulation. Multiple sequence alignment and conservation analysis of Human SFRPs showed that the CRD domain of SFRPs is more conserved than the NTR domain. The docking result indicated that SFRP3 has the highest binding affinity to Wnt3, while SFRP1 and SFRP5 have the lowest. Despite the lower affinity of SFRP1/SFRP5 for Wnt3, a higher positive charge in their NTR domains leads to an increase in their local concentration near the secreting cells and an enhancement in the antagonistic activity. In contrast, SFRP3/SFRP4 can act as an antagonist in distant cells due to less positive regions in their NTR domain and weakly binding to the heparin of the intercellular matrix.