Relationship of protein phosphorylation to the activation of the respiratory burst in human neutrophils. Defects in the phosphorylation of a group of closely related 48-kDa proteins in two forms of chronic granulomatous disease.
Relationship of protein phosphorylation to the activation of the respiratory burst in human neutrophils. Defects in the phosphorylation of a group of closely related 48-kDa proteins in two forms of chronic granulomatous disease.
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DOI：
10.1016/s0021-9258(18)68710-1
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发表时间：
1988-05
期刊：
影响因子：
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通讯作者：
N. Okamura;J. Curnutte;R. Roberts;B. Babior
中科院分区：
文献类型：
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作者：
N. Okamura;J. Curnutte;R. Roberts;B. Babior
当32 P标记的人中性粒细胞通过暴露于佛波醇肉豆蔻酸酯乙酸酯而被激活时，三种48-kDa蛋白（命名为pp 48/6.8，pp 4817. 3和pp 48/7.8）被标记。在最大刺激下，标记在30秒内完成。在较小程度的刺激下，2分钟时的标记程度与佛波醇处理的细胞的生产相关。在暴露于f-Met-Leu-Phe的细胞中也观察到这些48-kDa蛋白的标记增加。在X连锁细胞色素bss阴性慢性肉芽肿病患者的佛波醇治疗中性粒细胞中，pp 4817。pp 48/6.8和pp 48/7.3不能摄取~（32）P。然而，在常染色体隐性遗传的细胞色素bess阳性慢性肉芽肿病患者的细胞中，这三种蛋白质都没有对佛波醇产生反应。这三种蛋白质似乎是非常密切相关的，如磷酸丝氨酸是在任何三个中发现的唯一磷酸氨基酸的研究结果所示，所有三个产生相同的一维磷酸肽图消化后，无论是胰凝乳蛋白酶或葡萄球菌蛋白酶V8。这些结果与早期慢性肉芽肿病中蛋白磷酸化的观察结果相一致，并为这组48-kDa蛋白的磷酸化与呼吸爆发氧化酶的激活之间的关系提供了进一步的证据。“呼吸爆发”是一种代谢事件，在此期间，中性粒细胞和其他专职吞噬细胞通过部分氧气还原产生强大的抗微生物氧化剂（1）。所有这些抗微生物氧化剂的前体是超氧化物（O2），其通过以NADPH为代价的氧的单电子还原在活化的吞噬细胞中产生。
When 32P-labeled human neutrophils were activated by exposure to phorbol myristate acetate, three 48-kDa proteins (designated pp48/6.8, pp4817. 3, and pp48/7.8, from their isoelectric points) were found to have become labeled. With maximal stimulation, labeling was complete by 30 s. With lesser degrees of stimulation, the extent of labeling at 2 min correlated with of production by the phorbol-treated cells. Increased labeling of these 48-kDa proteins was also seen in cells exposed to f-Met-Leu-Phe. In phorbol-treated neutrophils from patients with X-linked cytochrome bsss-negative chronic granulomatous disease, pp4817. 8 was labeled in a normal fashion, but pp48/6.8 and pp48/7.3 failed to take up 32P. In cells from patients with autosomal recessive cytochrome bess-positive chronic granulomatous disease, however, none of the three proteins took up” P in response to phorbol. The three proteins appear to be very closely related, as indicated by the findings that phosphoserine was the only phosphoamino acid found in any of the three, and all three yielded identical one-dimensional phosphopeptide maps after digestion with either chymotrypsin or staphylococcal proteinase V8. These results reconcile earlier observations on protein phosphorylation in chronic granulomatous disease and provide further evidence for a relationship between the phosphorylation of this group of 48-kDa proteins and the activation of the respiratory burst oxidase.The “respiratory burst” is a metabolic event during which neutrophils and other professional phagocytes generate powerful antimicrobial oxidants by the partial reduction of oxygen (1). The precursor of all these antimicrobial oxidizing agents is superoxide (O;), which is generated in activated phagocytes by the one-electron reduction of oxygen at the expense of NADPH.