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. 2022 Sep 7;8(36):eabn0047. doi: 10.1126/sciadv.abn0047

Fig. 1. OBOC assay for human sperm binding and FcRL3 ERC with gamete recognition proteins.

Fig. 1.

(A) OBOC library structure (c, d-cysteine; X, 19 l-eukaryotic amino acids except l-cysteine). (B) Sperm-OBOC assay. (C) Sperm-bead binding (inset: human egg). (D) Sperm-bead binding (higher magnification). (E) Peptide hits and proportions assigned to functions. (F) Sperm (five donors) consistently bound to four specific bead aliquots incubated under specific conditions (fig. S1). Bead 16 hit for FcRL3. (G) Sperm bound to resynthesized cAMWNEDc peptide–beads during incubation. (H) Sperm-bound bead (higher magnification, bottom left corner; representative sperm highlighted by green cross) and “naked” bead (*). (I) IZUMO1 on human sperm. (J) Acrosomal antigen 18.6 on human sperm (control). (K) Antibody inhibition of sperm binding to resynthesized beads (n = repetitions) (16). (L) Antibody inhibition of sperm fusion to zona-free hamster eggs (n = animals, each of 10 to 15 oocytes). (M) Gamete interaction proteins on sperm (IZUMO1, SPACA6, DCST1, TMEM95, DCST2, and FIMP) and egg (JUNO, FcRL1, and FcRL3) experienced similar gene evolutionary histories, as shown by ERC sequence analysis. Color intensity reflects the strength of ERC value for that pair of genes. NA, not available.