Two-dimensional crystal structures of protein kinase C-delta, its regulatory domain, and the enzyme complexed with myelin basic protein.
Two-dimensional crystal structures of protein kinase C-delta, its regulatory domain, and the enzyme complexed with myelin basic protein.
复制标题
蛋白激酶 C-δ 的二维晶体结构、其调节域以及与髓磷脂碱性蛋白复合的酶。
DOI：
10.1016/s0006-3495(02)75611-7
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发表时间：
2002
影响因子：
3.4
通讯作者：
Kretsinger,RobertH
中科院分区：
文献类型：
--
作者：
Solodukhin,AlexanderS;Caldwell,HeatherL;Sando,JulianneJ;Kretsinger,RobertH
蛋白激酶C（PKC）δ、其调节结构域（RDδ）和与底物髓鞘碱性蛋白复合的酶的二维晶体已在由磷脂酰胆碱：磷脂酰丝氨酸：二油酸甘油酯（45：50：5，摩尔比）组成的脂质单层上生长。图像已被重建到10- 10厘米的分辨率。三种蛋白质的晶胞均具有晶胞边a=B，晶胞内角γ= 60 o。RDδ的边长为33± 1 π，其重构为圆环形。PKCδ C1 b晶体结构的三维重建（Zhang et al.，1995）可以容纳在该二维投影中。完整的PKCδ在存在或不存在不可水解的ATP类似物AMP-P β的情况下具有46± 1 μ m的边长。它的重建具有类似的圆环形状，可以容纳C1 b域，但圆环之间的间距大于RDδ;在圆环之间可以看到一些额外的结构。PKCδ与髓鞘碱性蛋白的复合物，无论有无AMP-PKC，其边长均为43± 1 nm，结构清晰。这些结果表明，RDδ的C1结构域在二维晶体中紧密地堆积在膜平面上，单位细胞中存在单个PKCδ分子，并且其与髓鞘碱性蛋白的相互作用引起酶的构象和/或堆积的变化。
Two-dimensional crystals of protein kinase C (PKC)δ, its regulatory domain (RDδ), and the enzyme complexed with the substrate myelin basic protein have been grown on lipid monolayers composed of phosphatidylcholine: phosphatidylserine: diolein (45:50:5, molar ratio). Images have been reconstructed to 10-Å resolution. The unit cells of all three proteins have cell edges a=b and interedge angleγ=60o. RDδhas an edge length of 33±1Å, and its reconstruction is donut shaped. The three-dimensional reconstructions from the PKCδC1b crystal structure (Zhang et al., 1995) can be accommodated in this two-dimensional projection. Intact PKCδhas an edge length of 46±1Å in the presence or absence of a nonhydrolyzable ATP analog, AMP-PnP. Its reconstruction has a similar donut shape, which can accommodate the C1b domain, but the spacing between donuts is greater than that in RDδ; some additional structure is visible between the donuts. The complex of PKCδand myelin basic protein, with or without AMP-PnP, has an edge length of 43±1Å and a distinct structure. These results indicate that the C1 domains of RDδare tightly packed in the plane of the membrane in the two-dimensional crystals, that there is a single molecule of PKCδin the unit cell, and that its interaction with myelin basic protein induces a shift in conformation and/or packing of the enzyme.