Qi D F, Schatzman R C, Mazzei G J, Turner R S, Raynor R L, Liao S, Kuo J F
Biochem J. 1983 Aug 1;213(2):281-8. doi: 10.1042/bj2130281.
Effects of polyamines on various protein kinases were investigated. It was found that both phospholipid-sensitive Ca2+-dependent protein kinase and myosin light-chain kinase (a calmodulin-sensitive species of Ca2+-dependent protein kinase) were inhibited to different degrees by polyamines, with an approximate order of inhibitory potency of spermine = 1, 12-diaminododecane greater than spermidine = 1, 10-diaminodecane much greater than cadaverine = putrescine. Kinetic analysis revealed that spermine inhibited the phospholipid-sensitive enzyme non-competitively with respect to Ca2+ (Ki = 0.84 mM) and phosphatidylserine (Ki = 0.90 mM); it also inhibited myosin light-chain kinase non-competitively with respect to Ca2+ (Ki = 1.82 mM) and calmodulin (Ki = 2.73 mM). 1, 12-Diaminododecane, in comparison, inhibited the phospholipid-sensitive enzyme competitively with respect to Ca2+ (Ki = 0.45 mM) and phosphatidylserine (Ki = 0.50 mM); it also inhibited myosin light-chain kinase competitively with respect to calmodulin (Ki = 0.63 mM) but non-competitively with respect to Ca2+ (Ki = 1.49 mM). Moreover, spermine (0.5 mM) was found to inhibit markedly phosphatidylserine/Ca2+- and calmodulin/Ca2+-stimulated phosphorylation of endogenous proteins in rat brain particulate fraction. All the polyamines tested were practically without effect on cyclic AMP-dependent and cyclic GMP-dependent protein kinases. Polyarginine, like spermine, was found to be a more selective inhibitor of Ca2+-dependent protein kinases, whereas polyglutamate preferentially inhibited the cyclic nucleotide-dependent enzymes. The present results indicated that, in addition to certain lipophilic compounds (such as trifluoperazine, palmitoylcarnitine, adriamycin and naphthalenesulphonamide) and polypeptides with hydrophobic regions (such as melittin and polymyxin B) previously reported, polycationic compounds (exemplified by polyamines) could also inhibit the two classes of Ca2+-dependent protein kinases requiring either phospholipid or calmodulin as a cofactor. Because of the high cellular concentration (up to 10 mM) and the differential effects of polyamines, it is suggested that spermine, and to smaller extents spermidine and putrescine, may be involved in the regulation of certain Ca2+-dependent protein-phosphorylation systems in vivo.
研究了多胺对各种蛋白激酶的作用。发现磷脂敏感的钙依赖性蛋白激酶和肌球蛋白轻链激酶（一种钙调蛋白敏感的钙依赖性蛋白激酶）均受到多胺不同程度的抑制，抑制效力的大致顺序为：精胺 = 1,12 - 二氨基十二烷 ＞ 亚精胺 = 1,10 - 二氨基癸烷 ＞ 尸胺 = 腐胺。动力学分析表明，精胺对磷脂敏感酶的抑制作用相对于Ca²⁺（Ki = 0.84 mM）和磷脂酰丝氨酸（Ki = 0.90 mM）是非竞争性的；它对肌球蛋白轻链激酶相对于Ca²⁺（Ki = 1.82 mM）和钙调蛋白（Ki = 2.73 mM）也是非竞争性抑制。相比之下，1,12 - 二氨基十二烷对磷脂敏感酶相对于Ca²⁺（Ki = 0.45 mM）和磷脂酰丝氨酸（Ki = 0.50 mM）是竞争性抑制；它对肌球蛋白轻链激酶相对于钙调蛋白（Ki = 0.63 mM）是竞争性抑制，但相对于Ca²⁺（Ki = 1.49 mM）是非竞争性抑制。此外，发现精胺（0.5 mM）能显著抑制大鼠脑微粒体部分中磷脂酰丝氨酸/Ca²⁺和钙调蛋白/Ca²⁺刺激的内源性蛋白磷酸化。所有测试的多胺实际上对环磷酸腺苷依赖性和环磷酸鸟苷依赖性蛋白激酶均无作用。聚精氨酸与精胺一样，是钙依赖性蛋白激酶更具选择性的抑制剂，而聚谷氨酸优先抑制环核苷酸依赖性酶。目前的结果表明，除了先前报道的某些亲脂性化合物（如三氟拉嗪、棕榈酰肉碱、阿霉素和萘磺酰胺）以及具有疏水区域的多肽（如蜂毒肽和多粘菌素B）外，聚阳离子化合物（以多胺为例）也能抑制两类需要磷脂或钙调蛋白作为辅因子的钙依赖性蛋白激酶。由于细胞内多胺浓度较高（高达10 mM）且具有不同作用，提示精胺以及程度较小的亚精胺和腐胺可能参与体内某些钙依赖性蛋白磷酸化系统的调节。
