Use of automated smoothing and deconvolution procedures for the determination of human breast cancer estrogen and progesterone receptor isoforms, after high-performance size-exclusion chromatography.
Automated smoothing and deconvolution procedures were used to analyse complex chromatographic patterns of human breast cancer estrogen and progesterone receptors, obtained by size-exclusion chromatography. By injecting, first, different known amounts of a radio-iodinated protein in a TSK G-3000 SW column, constructed complex chromatograms (twelve chromatograms) with known peak positions were obtained, and were further treated by mathematical functions to determine a smooth-deconvolution strategy, which could be used with unknown chromatographic patterns. The determination of peak areas by a "curve-fit" program showed a good correlation with the amounts of protein injected (r = 0.91). Human breast cancer estrogen receptors (56 cases) and progesterone receptors (45 cases) were chromatographed in a TSK G-3000 SW column, and further analysed with the smoothing and deconvolution procedures: eight different estrogen receptor isoforms were detected with molecular masses ranging from 530,000 [Stoke's radius (Rs) = 7.7 nm] to 23,000 (Rs = 2.6 nm), and eight progesterone receptor isoforms were observed with masses ranging from 680,000 (Rs = 8.6 nm) to 50,000 (Rs = 3.1 nm). The dissociation effect of KCl (0 to 1 M) on receptor structure yielded different proportions of receptor isoforms, but did not modify their different peak positions.
Cren
H
H
Laboratoire d'Analyses Isotopiques, Centre François Baclesse, Caen, France.
Lechevrel
C
C
Roussel
G
G
Lemoisson
E
E
Goussard
J
J
D001943:Breast Neoplasms / Q000378:metabolism*; D002850:Chromatography, Gel; D003600:Cytosol / Q000737:chemistry; D005583:Fourier Analysis; D006801:Humans; D007202:Indicators and Reagents; D007536:Isomerism; D011189:Potassium Chloride; D011869:Radioligand Assay; D011960:Receptors, Estrogen / Q000032:analysis*; D011980:Receptors, Progesterone / Q000032:analysis*; D005719:gamma-Globulins / Q000032:analysis
D007202:Indicators and Reagents; D011960:Receptors, Estrogen; D011980:Receptors, Progesterone; D005719:gamma-Globulins; D011189:Potassium Chloride