Identification of a putative inhibitor of early pregnancy factor in mice

Cavanagh, Alice; Morton, H. Craig; Athanasas‐Platsis, Stavrosia; Quinn, Kathryn; Rolfe, Barbara E.

doi:10.1530/jrf.0.0910239

Cited by 15 publications

(17 citation statements)

References 15 publications

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“…Ovaries from 50 superovulated mice were cultured in RPMI-1640 (ICN-Row) with the addition of rat prolactin (PRL-6 ; National Institutes of Health, Bethesda, USA) and medium conditioned by mouse embryos, as described previously (Cavanagh, 1984;Cavanagh et al, 1991b). A control culture was prepared with ovaries from 50 superovulated mice, cultured in RPMI-1640 medium alone; prolactin and mouse embryo-conditioned medium were added at the time of harvest.…”

Section: Mouse Ovary-conditioned Mediummentioning

confidence: 99%

The purification of early‐pregnancy factor to homogeneity from human platelets and identification as chaperonin 10

Cavanagh

Morton

1994

European Journal of Biochemistry

136

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Early‐pregnancy factor (EPF), first discovered in the early stages of gestation, is associated with and necessary for cell proliferation in a wide variety of biological situations. Like many other growth factors, EPF is present in platelets, and, by titration studies with a neutralising anti‐EPF monoclonal antibody, platelets were identified as an extremely rich source of this growth factor. EPF has been purified from clinically outdated human platelets by heat extraction, ion‐exchange and affinity chromatographies on SP‐Sephadex and heparin‐Sepharose respectively, high‐performance hydrophobic interaction chromatography and three reverse‐phase HPLC steps, with an average yield of 15 μg/100 platelet units (equivalent to ∼ 50 1 blood). Using SDS/PAGE, EPF migrated as a single band with approximate Mr 8500, coincident with biological activity. Mass spectrometry provided an accurate and precise determination of the molecular mass as Mr 10843.5±2, along with definitive evidence of the homogeneity of the preparation. Attempts at Edman degradation indicated that the molecule was blocked at the N‐terminus and sequencing of proteolytic fragments was undertaken. The amino acid sequence of approximately 70% of the molecule was determined which, with a single exception, is identical with rat chaperonin 10. This structural relationship was shown to extend to functional identity by studies using chaperonin 10 and its functional associate chaperonin 60. Investigations with the latter confirmed that chaperonin 10 is the moiety in pregnancy serum which initiates response in the EPF bioassay. Our studies identify EPF as a member of the highly conserved heat‐shock family of molecules and demonstrate a molecular chaperone performing an extracellular role.

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Section: Mouse Ovary-conditioned Mediummentioning

confidence: 99%

The purification of early‐pregnancy factor to homogeneity from human platelets and identification as chaperonin 10

Cavanagh

Morton

1994

European Journal of Biochemistry

136

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“…We looked for possible protocol differences either in the assay procedures or in the preparation of the samples to explain these differences. Compared with the assay procedure used by Cavanagh et al (1991) we used (i) a different lysis procedure in the preparation of the spleen cells; (ii) human serum as a source of complement not guinea-pig serum and (iii) the cell incubation step was performed in PBS not HBSS. Although these modifications of the assay have been docu¬ mented (Orozco et al, 1986;Clarke et al, 1987) .…”

Section: Introductionmentioning

confidence: 99%

The 'early pregnancy factor' revisited: the effect of ammonium sulfate on the capacity of pregnant mouse sera to express activity in the rosette inhibition assay

Clark

Orozco²,

Cock³

et al. 1994

Reproduction

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“…Bose et al [10] purified human EPF from the culture medium of preimplantation embryos and reported that EPF had a molecular weight of 14 KD. Smart [5] and other researchers [9][10][11][12] reported that EPF was released from human ova fertilized both in vitro and in vivo, but Morton et al [13], Chen [14], Nahhas et al [15], and Cavanagh et al [16] Bovine (Holstein) ova were obtained by aspirating ovarian follicles (<5 mm) within 4 h of slaughter. After the ova were cultured for 20 h in TCM-199 medium containing 5% fetal calf serum (FCS), they were fertilized in vitro (fertility rate = 50%).…”

mentioning

confidence: 99%

Study of EPF-Like Substance(s) Detected in Fertilized Bovine Ovum Culture Medium.

Ikemizu

Ito

Kawahata³

et al. 1994

Journal of Reproduction and Development

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Abstract. This study was undertaken to confirm whether early pregnancy factor (EPF)-like substance (s) was secreted by fertilized bovine ova. Using the rosette inhibition test, EPF-like substance(s) was detected in the culture media of fertilized bovine ova, while this substance was not found in the culture media of unfertilized bovine ova. EPF-like activity, analyzed by HPLC-gel permeation chromatography, was present between molecular weights of 16 and 37 KD. In conclusion, EPF-like substance(s) is secreted by fertilized bovine ovum and has a molecular weight of 16-37 KD.

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Identification of a putative inhibitor of early pregnancy factor in mice

Abstract: Summary. Previous

Cited by 15 publications

References 15 publications

The purification of early‐pregnancy factor to homogeneity from human platelets and identification as chaperonin 10

The purification of early‐pregnancy factor to homogeneity from human platelets and identification as chaperonin 10

The 'early pregnancy factor' revisited: the effect of ammonium sulfate on the capacity of pregnant mouse sera to express activity in the rosette inhibition assay

Study of EPF-Like Substance(s) Detected in Fertilized Bovine Ovum Culture Medium.

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