AMP PNP (adenylyl-imidodiphosphate) is a non-hydrolyzable analog of ATP. It can inhibit fast axonal transport and promote interactions between membranous organelles and microtubules [1]. AMP-PNP acts as a competitive inhibitor for most ATP-dependent systems, inhibiting a variety of ATP-hydrolyzing enzymes such as DNA topoisomerase II, SV40 large T antigen helicase, and several kinases. It also blocks ATP-sensitive, calcium-dependent potassium channels [2-4]. AMP-PNP can be used for the preparation of nucleotide solutions [5], dual-polarization fluorescence detection, actin cytoskeleton preparation for flow cells [6], and cascade binding reactions [7]. However, AMP-PNP is highly unstable under acidic conditions and rapidly hydrolyzes into phosphamide and inorganic phosphate.
References:
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[2]. Hehl S, Neumcke B. KATP channels of mouse skeletal muscle: mechanism of channel blockage by AMP-PNP. Eur Biophys J. 1994;23(4):231-7. doi: 10.1007/BF00213573. PMID: 7805625.
[3]. Subramanian R, Gelles J. Two distinct modes of processive kinesin movement in mixtures of ATP and AMP-PNP. J Gen Physiol. 2007 Nov;130(5):445-55. doi: 10.1085/jgp.200709866. PMID: 17968024; PMCID: PMC2151671.
[4]. Lasek RJ, Brady ST. Attachment of transported vesicles to microtubules in axoplasm is facilitated by AMP-PNP. Nature. 1985 Aug 15-21;316(6029):645-7. doi: 10.1038/316645a0. PMID: 4033761.
[5]. Fendley GA, Urbatsch IL, et,al. Nucleotide dependence of the dimerization of ATP binding cassette nucleotide binding domains. Biochem Biophys Res Commun. 2016 Nov 11;480(2):268-272. doi: 10.1016/j.bbrc.2016.10.046. Epub 2016 Oct 17. PMID: 27765627.
[6]. DeBerg HA, Blehm BH, et,al. Motor domain phosphorylation modulates kinesin-1 transport. J Biol Chem. 2013 Nov 8;288(45):32612-32621. doi: 10.1074/jbc.M113.515510. Epub 2013 Sep 26. PMID: 24072715; PMCID: PMC3820893.
[7]. Jung C, Hawkins JA, et,al. Massively Parallel Biophysical Analysis of CRISPR-Cas Complexes on Next Generation Sequencing Chips. Cell. 2017 Jun 29;170(1):35-47.e13. doi: 10.1016/j.cell.2017.05.044. PMID: 28666121; PMCID: PMC5552236.
AMP PNP (adenylyl-imidodiphosphate)是ATP的非水解类似物。它可抑制快速轴突转运，促进膜性细胞器和微管之间的相互作用[1]。AMP-PNP是大多数ATP性依赖系统的竞争性抑制剂，它可抑制一系列需要水解ATP的酶，比如DNA拓扑异构酶Ⅱ，SV40大T抗原螺旋酶和一系列激酶。阻断ATP敏感钙依赖性钾离子通道[2-4]。AMP-PNP可用于制备核苷酸溶液[5]，双动力荧光检测，结合肌动蛋白制备流动细胞[6]以及级联结合反应[7]。但是AMP-PNP在酸性条件下非常不稳定，会快速水解成磷酸酰胺和无机磷酸盐。