Methionine aminopeptidase 2 is a new target for the metastasis-associated protein, S100A4.
S100A4 is an EF-hand type calcium-binding protein that regulates tumor metastasis and a variety of cellular processes via interaction with different target proteins. Here we report that S100A4 physically interacts with methionine aminopeptidase 2 (MetAP2), the primary target for potent angiogenesis inhibitors, fumagillin and ovalicin. Using a yeast two-hybrid screen, S100A4 was found to interact with the N-terminal half of MetAP2. In vitro pull-down assays showed that S100A4 associates with MetAP2 in a calcium-dependent manner. In addition, the binding site of S100A4 was found located within the region between amino acid residues 170 and 229 of MetAP2. In vivo interaction of S100A4 with MetAP2 was verified by co-immunoprecipitation analysis. Immunofluorescent staining revealed that S100A4 and MetAP2 were co-localized in both quiescent and basic fibroblast growth factor-treated murine endothelial MSS31 cells, in the latter of which a significant change of intracellular distribution of both proteins was observed. Although the binding of S100A4 did not affect the in vitro methionine aminopeptidase activity of MetAP2, the cytochemical observation suggests a possible involvement of S100A4 in the regulation of MetAP2 activity through changing its localization, thereby modulating the N-terminal methionine processing of nascent substrates. These results may offer an essential clue for understanding the functional role of S100A4 in regulating endothelial cell growth and tumor metastasis.
Endo
Hideya
H
Division of Pathology, Department of Cancer Biology, The Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan. endoh@ims.u-tokyo.ac.jp
Takenaga
Keizo
K
Kanno
Takayuki
T
Satoh
Hitoshi
H
Mori
Shigeo
S
D000595:Amino Acid Sequence; D000626:Aminopeptidases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D020533:Angiogenesis Inhibitors / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002147:Calmodulin / Q000378:metabolism; D003510:Cyclohexanes; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism; D005231:Fatty Acids, Unsaturated / Q000494:pharmacology; D005346:Fibroblast Growth Factors / Q000494:pharmacology; D006801:Humans; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D008715:Methionine / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D009362:Neoplasm Metastasis; D000071999:S100 Calcium-Binding Protein A4; D009418:S100 Proteins / Q000378:metabolism*; D012717:Sesquiterpenes / Q000494:pharmacology; D018411:Spodoptera; D014407:Tumor Cells, Cultured
D020533:Angiogenesis Inhibitors; D002147:Calmodulin; D003510:Cyclohexanes; D005231:Fatty Acids, Unsaturated; D000071999:S100 Calcium-Binding Protein A4; D009418:S100 Proteins; C470944:S100a4 protein, mouse; D012717:Sesquiterpenes; C077591:S100A4 protein, human; C002781:ovalicin; D005346:Fibroblast Growth Factors; C026211:fumagillin; D008715:Methionine; D000626:Aminopeptidases; C106541:methionine aminopeptidase 2; D008666:Metalloendopeptidases; D002118:Calcium