PUBLICATION
Identification of a residue crucial for the angiostatic activity of human mini tryptophanyl-tRNA synthetase by focusing on its molecular evolution
- Authors
- Nakamoto, T., Miyanokoshi, M., Tanaka, T., Wakasugi, K.
- ID
- ZDB-PUB-160421-12
- Date
- 2016
- Source
- Scientific Reports 6: 24750 (Journal)
- Registered Authors
- Wakasugi, Keisuke
- Keywords
- RNA-binding proteins
- MeSH Terms
-
- Amino Acid Substitution
- Amino Acids*
- Aminoacylation
- Animals
- Binding Sites
- Cadherins/metabolism
- Chemotaxis
- Endothelial Cells/metabolism
- Enzyme Activation
- Evolution, Molecular*
- Humans
- Protein Binding
- Tryptophan-tRNA Ligase/chemistry*
- Tryptophan-tRNA Ligase/genetics*
- Tryptophan-tRNA Ligase/metabolism
- Vascular Endothelial Growth Factor A/metabolism
- PubMed
- 27094087 Full text @ Sci. Rep.
Citation
Nakamoto, T., Miyanokoshi, M., Tanaka, T., Wakasugi, K. (2016) Identification of a residue crucial for the angiostatic activity of human mini tryptophanyl-tRNA synthetase by focusing on its molecular evolution. Scientific Reports. 6:24750.
Abstract
Human tryptophanyl-tRNA synthetase (TrpRS) exists in two forms: a full-length TrpRS and a mini TrpRS. We previously found that human mini, but not full-length, TrpRS is an angiostatic factor. Moreover, it was shown that the interaction between mini TrpRS and the extracellular domain of vascular endothelial (VE)-cadherin is crucial for its angiostatic activity. However, the molecular mechanism of the angiostatic activity of human mini TrpRS is only partly understood. In the present study, we investigated the effects of truncated (mini) form of TrpRS proteins from human, bovine, or zebrafish on vascular endothelial growth factor (VEGF)-stimulated chemotaxis of human umbilical vein endothelial cells (HUVECs). We show that both human and bovine mini TrpRSs inhibited VEGF-induced endothelial migration, whereas zebrafish mini TrpRS did not. Next, to identify residues crucial for the angiostatic activity of human mini TrpRS, we prepared several site-directed mutants based on amino acid sequence alignments among TrpRSs from various species and demonstrated that a human mini K153Q TrpRS mutant cannot inhibit VEGF-stimulated HUVEC migration and cannot bind to the extracellular domain of VE-cadherin. Taken together, we conclude that the Lys153 residue of human mini TrpRS is a VE-cadherin binding site and is therefore crucial for its angiostatic activity.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping