Combinatorial peptide library binding of mammalian spermatozoa identifies a ligand (HIPRT) in the axin protein: putative identification of a sperm surface axin binding protein and intriguing developmental implications.
The identification of components in cell-cell interactions is an important research goal in reproductive and developmental biology. Such interactions are critical to gamete development, fertilization, implantation and basic development. Several proteins involved with sperm-oocyte interaction and other developmentally important phenomena have been identified. However, these are obviously only a subset of the molecular components involved in such complex cell-cell interactions. One method that has been used to identify binding partners for particular molecular targets is the use of combinatorial libraries accessible on phage surfaces. For the most part, this technique has mainly been applied to screen specific target moieties. However, in some cases whole-cell screening has been attempted. This study describes the first report of screening intact, living mammalian gametes using a proprietary whole-cell combinatorial library binding and analysis protocol. Results from the first screening protocol of mouse spermatozoa strongly identified a putative sperm-binding ligand using proprietary bioinformatic analysis. This amino acid sequence (HIPRT) precisely corresponds with a previously characterized highly conserved protein-protein interaction site in the axin protein. This sequence is found within the binding site for a known sperm surface protein, glycogen synthase kinase-3. This result not only provides proof of the utility of this technique to identify cell surface ligands in mammalian gametes, but it also suggests a potential role for spermatozoa in facilitating developmental axis formation in mammalian embryos.
Malter
Henry E
HE
Tyho-Galileo Research Laboratories, 101 Old Short Hills Road, Suite 501, West Orange, NJ 07052, USA.
Cohen
Jacques
J
Pieczenik
George
G
D000595:Amino Acid Sequence; D000818:Animals; D060466:Axin Protein; D001665:Binding Sites; D002352:Carrier Proteins / Q000302:isolation & purification / Q000378:metabolism*; D038362:Glycogen Synthase Kinase 3 / Q000737:chemistry / Q000378:metabolism; D066298:In Vitro Techniques; D008024:Ligands; D008297:Male; D008565:Membrane Proteins / Q000302:isolation & purification / Q000378:metabolism*; D051379:Mice; D019151:Peptide Library; D012097:Repressor Proteins / Q000737:chemistry / Q000378:metabolism*; D013094:Spermatozoa / Q000378:metabolism*
D060466:Axin Protein; D002352:Carrier Proteins; D008024:Ligands; D008565:Membrane Proteins; D019151:Peptide Library; D012097:Repressor Proteins; D038362:Glycogen Synthase Kinase 3