A5089920828- Sperm and Testicular Function
- Reproductive Biology and Fertility
- Animal Genetics and Reproduction
- Plant Reproductive Biology
- Monoclonal and Polyclonal Antibodies Research
- RNA Interference and Gene Delivery
- Protein Structure and Dynamics
- Protein purification and stability
- DNA and Nucleic Acid Chemistry
- RNA and protein synthesis mechanisms
- Cytomegalovirus and herpesvirus research
- Phytase and its Applications
- Enzyme Structure and Function
- Reproductive System and Pregnancy
- Antimicrobial Peptides and Activities
- Polyamine Metabolism and Applications
- Immunodeficiency and Autoimmune Disorders
Karolinska Institutet
2016-2025
Recognition between sperm and the egg surface marks beginning of life in all sexually reproducing organisms. This fundamental biological event depends on species-specific interaction rapidly evolving counterpart molecules gametes. We report biochemical, crystallographic, mutational studies domain repeats 1-3 invertebrate coat protein VERL their with cognate lysin. fold like functionally essential N-terminal repeat mammalian receptor ZP2, whose structure is also described here. Whereas...
Following the fertilization of an egg by a single sperm, coat or zona pellucida (ZP) hardens and polyspermy is irreversibly blocked. These events are associated with cleavage N-terminal region (NTR) glycoprotein ZP2, major subunit ZP filaments. ZP2 processing thought to inactivate sperm binding ZP, but its molecular consequences connection hardening unknown. Biochemical structural studies show that triggers oligomerization. Moreover, structure native vertebrate filament, combined AlphaFold...
Abstract Mammalian fertilisation begins when sperm interacts with the egg zona pellucida (ZP), whose ZP1 subunit is important for fertility by covalently cross-linking ZP filaments into a three-dimensional matrix. Like ZP4, structurally-related component absent in mouse, predicted to contain an N-terminal ZP-N domain of unknown function. Here we report characterisation proteins carrying mutations from infertile patients, which suggests that, human, filament crucial form stable ZP. We map...
We present a strategy to obtain milligrams of highly post-translationally modified eukaryotic proteins, transiently expressed in mammalian cells as rigid or cleavable fusions with mammalianized version bacterial maltose-binding protein (mMBP). This variant was engineered combine mutations that enhance MBP solubility and affinity purification, well provide crystal-packing interactions for increased crystallizability. Using this cell type-independent approach, we could increase the expression...
Monoclonal antibody IE-3 prevents mouse fertilization by binding ZP2, a major component of the oocyte-specific zona pellucida (ZP). We show that an IE3-derived single-chain variable fragment (scFV) is sufficient for blocking in vitro and determine structural basis IE-3/ZP2 recognition. The high-affinity this interaction depends on induced fit epitope, offering insights non-hormonal immunocontraceptive design without off-target effects.
Monoclonal antibody IE-3 prevents mouse fertilization by binding ZP2, a major component of the oocyte-specific zona pellucida (ZP). We show that an IE-3-derived single-chain variable fragment (scFV) is sufficient for blocking in vitro and determine structural basis IE-3/ZP2 recognition. The high affinity this interaction depends on induced fit epitope, offering insights nonhormonal contraceptive design without off-target effects.
Abstract Interaction between sperm and the egg zona pellucida (ZP) is first step of mammalian fertilization, ZP component ZP1 important for fertility by covalently cross-linking filaments into a matrix. Like ZP4, structurally-related subunit absent in mouse, predicted to contain an N-terminal ZP-N domain unknown function. Characterization proteins carrying mutations from infertile patients suggests that, unlike filament crucial human assembly. We map function its ZP-N1 determine crystal...
SUMMARY Post-fertilization cleavage of glycoprotein ZP2, a major subunit egg zona pellucida (ZP) filaments, is crucial for mammalian reproduction by irreversibly blocking polyspermy. ZP2 processing thought to inactivate sperm-binding activity located upstream the protein’s site; however, its molecular consequences and connection with ZP hardening are unknown. Here we report X-ray crystallographic, cryo-EM biochemical studies showing that triggers oligomerization. Deletion ZP-N1 domain...
Post-fertilization cleavage of glycoprotein ZP2, a major subunit egg zona pellucida (ZP) filaments, is crucial for mammalian reproduction by irreversibly blocking polyspermy. ZP2 processing thought to inactivate sperm-binding activity located upstream the protein’s site; however, its molecular consequences and connection with ZP hardening are unknown. Here we report X-ray crystallographic, cryo-EM biochemical studies showing that triggers oligomerization. Deletion ZP-N1 domain precedes site...