Kirkness E F, Turner A J
Department of Biochemistry, University of Leeds, U.K.
Biochem J. 1988 Nov 15;256(1):291-4. doi: 10.1042/bj2560291.
A synthetic peptide, corresponding to amino acid residues 101-109 of the bovine gamma-aminobutyrate/benzodiazepine receptor alpha-subunit, was used to raise a polyclonal antiserum. The reactivity of this antiserum towards polypeptides of both bovine and pig receptor preparations was established by immunoprecipitation and immunoblotting. Anti-peptide antibodies recognized the alpha-subunit (51 kDa) of receptor prepared from pig cerebellum or cerebral cortex. However, a polypeptide of 57 kDa was additionally recognized in cortical, but not cerebellar, preparations. This alpha-like polypeptide appeared larger than the band of polypeptides labelled irreversibly with [3H]muscimol (beta-subunit, 55-57 kDa) and corresponds to a polypeptide detected only in cortex after silver-staining or irreversible labelling with [3H]flunitrazepam. These results support the idea that the distinct regional patterns of polypeptides labelled irreversibly with [3H]flunitrazepam reflect the existence of heterologous distributions of distinct alpha-like subunits.
一种对应于牛γ-氨基丁酸/苯二氮䓬受体α亚基第101 - 109位氨基酸残基的合成肽被用于制备多克隆抗血清。通过免疫沉淀和免疫印迹确定了该抗血清对牛和猪受体制剂中多肽的反应性。抗肽抗体识别了从猪小脑或大脑皮质制备的受体的α亚基(51 kDa)。然而,在皮质制剂而非小脑组织中还识别出了一条57 kDa的多肽。这条α样多肽看起来比用[³H]蝇蕈醇不可逆标记的多肽条带(β亚基,55 - 57 kDa)更大,并且与仅在皮质中经银染或用[³H]氟硝西泮不可逆标记后检测到的一条多肽相对应。这些结果支持了这样一种观点,即[³H]氟硝西泮不可逆标记的多肽的不同区域模式反映了不同α样亚基的异源分布的存在。