Wiehle R D, Hofmann G E, Fuchs A, Wittliff J L
J Chromatogr. 1984 Apr 13;307(1):39-51. doi: 10.1016/s0378-4347(00)84071-2.
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 greater than 61 A, approximately 48 A and 29-32 A. 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-17 beta 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.
采用TSK 3000SW分子筛柱进行高效尺寸排阻色谱法（HPSEC），基于大小和形状（斯托克斯半径）在50分钟内从多个靶组织中分离雌激素、孕激素和雄激素受体。此外，该系统检测受体种类（异构体）异质性的方式优于蔗糖密度梯度离心法。HPSEC分离出了斯托克斯半径大于61 Å、约48 Å以及29 - 32 Å的各种雌激素受体异构体。诸如氯化钾和钼酸钠等试剂，在蔗糖密度梯度离心时会改变雌激素受体种类的分布，采用HPSEC时也会促使受体图谱发生类似变化。我们的研究表明，使用[125I]碘雌二醇 - 17β和HPSEC可对雌激素受体种类进行顺序分析，从而为受体组成和结构提供新的见解。得出的结论是，HPSEC在类固醇激素受体领域具有广泛应用。该方法在从分子和动力学性质测量到细胞相互作用和调节模式的研究中应会很有用。
