PUBLICATION
Crystal Structure of Zebrafish Hatching Enzyme 1 from the Zebrafish Danio rerio
- Authors
- Okada, A., Sano, K., Nagata, K., Yasumasu, S., Ohtsuka, J., Yamamura, A., Kubota, K., Iuchi, I., and Tanokura, M.
- ID
- ZDB-PUB-100826-6
- Date
- 2010
- Source
- Journal of molecular biology 402(5): 865-878 (Journal)
- Registered Authors
- Keywords
- metalloprotease, zebrafish, zinc protease, crystal structure, hatching enzyme
- MeSH Terms
-
- Mutagenesis, Site-Directed
- Protein Conformation
- Animals
- Zebrafish*
- Amino Acid Sequence
- Sequence Alignment
- Catalytic Domain
- Molecular Sequence Data
- Crystallography, X-Ray
- Metalloendopeptidases/chemistry*
- Metalloendopeptidases/metabolism*
- Protein Structure, Tertiary
- Amino Acid Substitution
- Models, Molecular
- PubMed
- 20727360 Full text @ J. Mol. Biol.
Citation
Okada, A., Sano, K., Nagata, K., Yasumasu, S., Ohtsuka, J., Yamamura, A., Kubota, K., Iuchi, I., and Tanokura, M. (2010) Crystal Structure of Zebrafish Hatching Enzyme 1 from the Zebrafish Danio rerio. Journal of molecular biology. 402(5):865-878.
Abstract
Fish hatching enzymes are zinc metalloproteases that digest the egg envelope (chorion) at the time of hatching. The crystal structure of the zebrafish hatching enzyme 1 (ZHE1) has been solved at 1.10-A resolution. ZHE1 is monomeric, mitten-shaped, and has a cleft at the center of the molecule. ZHE1 consists of three 3(10)-helices, three alpha-helices, and two beta-sheets. The central cleft represents the active site of the enzyme that is crucial for substrate recognition and catalysis. Alanine-scanning mutagenesis of the two substrate peptides has shown that Asp(P1') contributes the most and the residues at P4-P2' also contribute to the recognition of the major substrate peptide by ZHE1, whereas Glu(P3') and the hydrophobic residues at P4-P2, P2' and P5' contribute significantly to the recognition of the minor substrate peptide by ZHE1. Molecular models of these two substrate peptides bound to ZHE1 have been built based on the crystal structure of a transition-state analog inhibitor bound to astacin. In the substrate recognition models, the Asp(P1') in the major substrate peptide forms a salt bridge with Arg(182) of ZHE1, while the Glu(P3') in the minor substrate peptide instead forms a salt bridge with Arg(182). Thus these two substrate peptides would be differently recognized by ZHE1. The shapes and electrostatic potentials of the substrate-binding clefts of ZHE1 and the structurally similar proteins, astacin and bone morphogenetic protein 1 (BMP1), are significantly dissimilar due to different side chains, which would confer their distinctive substrate preferences.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping