Decreased phosphorylation of a low molecular weight protein by cGMP-dependent protein kinase in variant HL-60 cells resistant to nitric oxide- and cGMP-induced differentiation.
Decreased phosphorylation of a low molecular weight protein by cGMP-dependent protein kinase in variant HL-60 cells resistant to nitric oxide- and cGMP-induced differentiation.
复制标题
在对一氧化氮和 cGMP 诱导的分化具有抵抗力的变体 HL-60 细胞中，cGMP 依赖性蛋白激酶降低了低分子量蛋白质的磷酸化。
DOI：
10.1023/a:1006834324419
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发表时间：
1998
影响因子：
4.3
通讯作者：
Boss,GR
中科院分区：
文献类型：
--
作者：
Scheele,JS;Pilz,RB;Clark,G;Gupta,N;Loo,D;Martis,P;Boss,GR
我们之前描述了分离的HL-60细胞的一个变异亚系，它不能对一氧化氮(NO)生成剂或cGMP类似物做出反应而分化[7]。变异细胞具有正常的鸟苷环化酶活性和正常的NO诱导的细胞内cGMP浓度增加。我们现在通过体外和体内实验证明变异细胞具有正常的cGMP依赖的蛋白激酶(G-激酶)活性，并使用双向凝胶电泳法在亲本细胞中鉴定了六种G-激酶底物。在这六种蛋白质中，我们发现在体外和在完整细胞中，变异细胞中的一种蛋白质的磷酸化程度明显低于亲本细胞，通过35S-蛋氨酸/35S-半胱氨酸掺入，我们在变异细胞中发现了比亲本细胞少得多的这种蛋白质。该蛋白是cAMP依赖的蛋白激酶(A-Kinase)的共同底物；由于cAMP类似物仍能诱导变异细胞分化，因此，在诱导HL-60细胞分化过程中，NO/cGMP/G-Kinase和cAMP/A-Kinase信号转导通路似乎共享部分但不是全部相同的靶蛋白。
We previously described the isolation of a variant subline of HL-60 cells that does not differentiate in response to nitric oxide (NO)-generating agents or to cGMP analogs [7]. The variant cells have normal guanylate cyclase activity and normal NO-induced increases in the intracellular cGMP concentration. We now show that the variant cells have normal cGMP-dependent protein kinase (G-kinase) activity, both by an in vitro and in vivo assay, and using two-dimensional gel electrophoresis we have identified six G-kinase substrates in the parental cells. Of these six proteins, we found considerably less phosphorylation of one of the proteins in the variant cells than in parental cells, both in vitro and in intact cells, and by35S-methionine/35S-cysteine incorporation we found much less of this protein in the variant cells than in parental cells. The protein is a shared substrate of cAMP-dependent protein kinase (A-kinase); since cAMP analogs still induce differentiation of the variant cells, it appears that the NO/cGMP/G-kinase and cAMP/A-kinase signal transduction pathways share some but not all of the same target proteins in inducing differentiation of HL-60 cells.