Lee M J, Goldsworthy G J
Department of Biology, Birkbeck College, University of London, England.
Arch Insect Biochem Physiol. 1998;39(1):9-17. doi: 10.1002/(SICI)1520-6327(1998)39:1<9::AID-ARCH3>3.0.CO;2-E.
N alpha-tosyl-L-lysine chloromethyl ketone (TLCK) stimulates lipid synthesis in locust fat body in vitro, and is able to reverse the inhibitory effects of AKH-I on lipid synthesis. Effective stimulatory concentrations of TLCK were in the range of 0.2-1.0 mM. Similar stimulatory effects were also achieved with phenylalanine chloromethyl ketone (PheCK) and leucine chloromethyl ketone (LeuCK), but not with tosyl-phenylalanine chloromethyl ketone (TPCK), dansyl-glu-gly-arg-CK, chloroacetone, chloroacetic acid, chloroacetamide, chloroacetaldehyde, chloroacetyl-L-leucine or acetylated or fluorescamine-labelled TLCK, PheCK, and LeuCK. The level of stimulation caused by TLCK was dependent on incubation time, so that after a 5-h preincubation of fat body tissue with TLCK the stimulated rate was severalfold higher than the control. TLCK also increased the rate of uptake of trehalose and uridine, but not glucose, deoxyglucose or glycine. Increasing concentrations of bovine serum albumin (BSA) in the incubation medium caused a reduction in the rate of TLCK-stimulated acetate uptake, such that levels of uptake were no higher with 1% BSA than in the controls. A range of more specific protease and kinase inhibitors was tested, but none caused stimulation; thus the mode of action of TLCK on the stimulation of acetate uptake has yet to be identified. Elucidation of the mode of action of TLCK may facilitate the development of novel compounds for insect pest control.
Nα-对甲苯磺酰-L-赖氨酸氯甲基酮（TLCK）在体外刺激蝗虫脂肪体中的脂质合成，并能够逆转促动素-I（AKH-I）对脂质合成的抑制作用。TLCK的有效刺激浓度范围为0.2-1.0 mM。苯丙氨酸氯甲基酮（PheCK）和亮氨酸氯甲基酮（LeuCK）也有类似的刺激作用，但对甲苯磺酰-苯丙氨酸氯甲基酮（TPCK）、丹磺酰-谷氨酰胺-甘氨酸-精氨酸氯甲基酮、氯丙酮、氯乙酸、氯乙酰胺、氯乙醛、氯乙酰-L-亮氨酸或乙酰化或荧光胺标记的TLCK、PheCK和LeuCK则没有。TLCK引起的刺激水平取决于孵育时间，因此脂肪体组织用TLCK预孵育5小时后，刺激速率比对照高出几倍。TLCK还提高了海藻糖和尿苷的摄取速率，但对葡萄糖、脱氧葡萄糖或甘氨酸没有影响。孵育培养基中牛血清白蛋白（BSA）浓度的增加导致TLCK刺激的乙酸摄取速率降低，因此1% BSA时的摄取水平并不比对照高。测试了一系列更特异的蛋白酶和激酶抑制剂，但均未引起刺激；因此，TLCK刺激乙酸摄取的作用方式尚未确定。阐明TLCK的作用方式可能有助于开发用于害虫防治的新型化合物。
