Abstract
High-performance size exclusion chromatography (HPSEC) on TSK 3000SW molecular sieve columns was used to separate estrogen, progestin, and androgen receptors from several target tissues within 50 min on the basis of size and shape (Stokes radius). Moreover, this system provided for the detection of heterogeneity of receptor species (isoforms) in a manner superior to that observed with sucrose density gradient centrifugation. HPSEC separated various estrogen receptor isoforms having Stokes radii of > 61 Å, ∼ 48 Å and 29–32 Å. Agents such as potassium chloride and sodium molybdate which alter the distribution of estrogen receptor species on sucrose density gradient centrifugation, promote similar alterations in receptor profiles when HPSEC was employed. Our investigations suggest the use of [ 125I]iodoestradiol-17β and HPSEC allows the sequential analysis of estrogen receptor species providing new insights into receptor composition and structure. It is concluded that HPSEC has a broad application in the field of steroid hormone receptors. This method should be useful in studies ranging from measurements of molecular and kinetic properties to their mode of cellular interaction and regulation.
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