Wangsgard W P, Dasgupta M, Blumenthal D K
Department of Pharmacology & Toxicology, University of Utah, Salt Lake City 84112, USA.
Biochem Biophys Res Commun. 1997 Jan 3;230(1):179-83. doi: 10.1006/bbrc.1996.5927.
The gamma-subunit of phosphorylase kinase contains a protein kinase catalytic domain (residues 20-276) and a regulatory domain (residues 276-386). The purpose of the present investigation was to develop monospecific antibodies against four synthetic gamma-subunit regulatory domain peptides (PhK1: 362-386; PhK5: 342-366; PhK9: 322-346; PhK13: 302-326) to use as probes to study the structure of the regulatory domain. Each affinity-purified antibody was characterized with regard to its ability to bind three different structural forms of the gamma-subunit: the isolated gamma-subunit, the gamma-delta complex, and the holoenzyme complex (alpha beta delta gamma)4. Of the four antibodies, binding of affinity-purified anti-PhK13 was most affected by alterations in gamma-subunit interactions. Taken together, the data from this investigation indicate that the regulatory domain of the gamma-subunit can assume different immunochemically distinguishable conformations as the result of interactions among the alpha-, beta-, gamma-, and delta-subunits of phosphorylase kinase.
磷酸化酶激酶的γ亚基包含一个蛋白激酶催化结构域（第20 - 276位氨基酸残基）和一个调节结构域（第276 - 386位氨基酸残基）。本研究的目的是制备针对四种合成的γ亚基调节结构域肽段（PhK1：362 - 386；PhK5：342 - 366；PhK9：322 - 346；PhK13：302 - 326）的单特异性抗体，用作研究调节结构域结构的探针。对每种亲和纯化抗体结合γ亚基三种不同结构形式的能力进行了表征：分离的γ亚基、γ - δ复合物和全酶复合物（αβδγ）4。在这四种抗体中，亲和纯化的抗PhK13的结合受γ亚基相互作用改变的影响最大。综合来看，本研究的数据表明，由于磷酸化酶激酶的α、β、γ和δ亚基之间的相互作用，γ亚基的调节结构域可以呈现出不同的免疫化学可区分构象。
