Honkanen R E, Zwiller J, Moore R E, Daily S L, Khatra B S, Dukelow M, Boynton A L
Cancer Research Center of Hawaii University of Hawaii, Manoa, Honolulu 96813.
J Biol Chem. 1990 Nov 15;265(32):19401-4.
The level of protein phosphorylation is dependent on the relative activities of both protein kinases and protein phosphatases. By comparison with protein kinases, however, there have been considerably fewer studies on the functions of serine/threonine protein phosphatases. This is partly due to a lack of specific protein phosphatase inhibitors that can be used as probes. In the present study we characterize the inhibitory effects of microcystin-LR, a hepatotoxic cyclic peptide associated with most strains of the blue-green algae Microcystis aeruginosa found in the Northern hemisphere, that proves to be a potent inhibitor of type 1 (IC50 = 1.7 nM) and type 2A (IC50 = 0.04 nM) protein phosphatases. Microcystin-LR inhibited the activity of both type 1 and type 2A phosphatases greater than 10-fold more potently than okadaic acid under the same conditions. Type 2A protein phosphatases in dilute mammalian cell extracts were found to be completely inhibited by 0.5 nM microcystin-LR while type 1 protein phosphatases were only slightly affected at this concentration. Thus, microcystin-LR may prove to be a useful probe for the study and identification cellular processes which are mediated by protein phosphatases.
蛋白质磷酸化水平取决于蛋白激酶和蛋白磷酸酶的相对活性。然而,与蛋白激酶相比,对丝氨酸/苏氨酸蛋白磷酸酶功能的研究要少得多。部分原因是缺乏可用作探针的特异性蛋白磷酸酶抑制剂。在本研究中,我们表征了微囊藻毒素-LR的抑制作用,微囊藻毒素-LR是一种与北半球发现的大多数铜绿微囊藻菌株相关的肝毒性环肽,它被证明是1型(IC50 = 1.7 nM)和2A型(IC50 = 0.04 nM)蛋白磷酸酶的有效抑制剂。在相同条件下,微囊藻毒素-LR抑制1型和2A型磷酸酶的活性比冈田酸强10倍以上。发现在稀释的哺乳动物细胞提取物中,0.5 nM微囊藻毒素-LR可完全抑制2A型蛋白磷酸酶,而在此浓度下1型蛋白磷酸酶仅受到轻微影响。因此,微囊藻毒素-LR可能被证明是研究和鉴定由蛋白磷酸酶介导的细胞过程的有用探针。