IDENTIFICATION AND TISSUE LOCALIZATION OF AN EOSINOPHIL 17-KDA PROTEIN ACCUMULATING IN RAT UTERUS UPON ESTRADIOL TREATMENT

In a previous paper (J. Steroid Biochem. 29 (1988) 475-480), the isolation of a 17 kDa protein that was dramatically induced in the uterus of estrogen-treated spayed rats was presented. We now describe a new purification procedure that is compatible with microsequencing of the 17 kDa protein. The protein partial N-terminal amino acid sequence analysis gave 28 residues that revealed a strong homology to the human major basic protein (MBP) of eosinophils described by Wasmoen et al. (J. Biol. Chem. 263 (1988) 12559-12563). Polyclonal rabbit antibodies were raised against this protein and used for tissue or blood cell analysis after electrophoresis and Western blotting. The 17 kDa protein was found to be constitutively present in the stomach and small intestine of the rat and guinea-pig. Estrogen treatment had a clearcut effect in guinea-pig uterus, but not as drastic as that observed in rat uterus. The protein was abundant in purified rat eosinophils. The antibodies cross-reacted with human MBP and an equivalent molecular weight human polymorphonuclear leukocyte protein. Immunohistochemical staining of rat uterus sections showed that the protein was first only associated with eosinophils that emigrate upon estrogen treatment; it then spread throughout the stroma and the deep glandular epithelium. It was not found in the myometrium. In conclusion, the appearance of a 17 kDa protein that is presumably the rat MBP is clearly regulated in the rat uterus.