蛋白激酶A RⅡβ结构及功能的研究进展

doi:10.16098/j.issn.0529-1356.2017.06.022

摘要/Abstract

摘要：

蛋白激酶A（PKA）是由1个调节亚基（R亚基）二聚体和两个催化亚基（C亚基）组成的四聚体，全酶无活性。R亚基有RⅠα、RⅠβ、RⅡα和RⅡβ 4种亚型，分别具有不同的理化性质。RⅡβ亚基氨基端包含1个二聚化/对接结构域，PKA通过此结构域与腺苷酸激酶锚定蛋白结合锚定于细胞的特定位点。羧基端是两个串联的高度保守的环核苷酸结构域，与PKA 全酶解聚以及激活有关。两个RC异二聚体通过RⅡβ亚基中的β4~β5环相锚定。细胞质中存在足够的MgATP时将导致 RⅡβ亚基自身磷酸化。RⅡβ在特定组织表达，主要表达于内分泌腺、脑和脂肪组织。生物信息学分析表明，RⅡβ与其他R亚基的序列有很大差别，只有大约50%的序列相同，提示RⅡβ可能具有不同的生物学功能。因此，目前对PKA RⅡβ的功能及其作用机制的研究已逐渐成为热点。

关键词: 蛋白激酶A, 调节亚基, 变构效应, 磷酸化

Abstract:

Protein kinase A (PKA) exists in mammalian cells as an inactive tetrameric holoenzyme composed of a regulatory (R) subunit dimer and two catalytic (C) subunits. There are four isoforms of the R subunits, RIα, RIβ, RⅡα,and RⅡβ. Each of them has special physicochemical property. RⅡβ subunit contains an N-terminal dimerization/docking domain. PKA is anchored to specific sites in the cell by binding of an adenosine kinase-anchoring protein amphipathic helix to the dimerization/docking domain. At the C terminus there are two tandem highly conserved cyclic nucleotide-binding domains, which relate to depolymerization and activation of the holoenzyme. The heterodimers are anchored together by an interface created by the β4-β5 loop in the RⅡβ subunit. In cells the inactive RⅡβ2:C2 holoenzyme once formed, it may be primed to have its R subunits autophosphory when high availability of cytoplasmic MgATP is given. RⅡβ expression is tissue specific，mainly in the brain, adipose tissue and endocrine organs. Bioinormatics analysis has showed that the sequence of RⅡβis significantly different from other three R subunit isoforms. The big difference indicates that PKA RⅡβ may have special biological function. Therefore many researchers focus on the function and mechanism of PKA RⅡβ.

Key words: Protein kinase A, Regulatory subunit, Allosteric effect, Phosphorylation

丁玉静金兴刘俊秀马芙蓉. 蛋白激酶A RⅡβ结构及功能的研究进展[J]. 解剖学报, 2017, 48(6): 761-765.

DING Yu-jing JIN Xing LIU Jun-xiu MA Fu-rong. Progress of the function and mechanism of protein kinase A RⅡβ[J]. Acta Anatomica Sinica, 2017, 48(6): 761-765.

参考文献

［1］Zhang P, Smith-Nguyen EV, Keshwani MM, et al. Structure and allostery of the PKA RII beta tetrameric holoenzyme［J］. Science,2012,335(6069):712-716.

［2］Kim C, Vigil D, Anand G, et al. Structure and dynamics of PKA signaling proteins［J］. Eur J Cell Biol,2006,85(7):651-654.

［3］Shemarova IV. cAMP-dependent signal pathways in unicellular eukaryotes［J］. Crit Rev Microbiol,2009,35(1):23-42.

［4］Wojtal KA, Hoekstra D, van Ijzendoorn SC. cAMP-dependent protein kinase A and the dynamics of epithelial cell surface domains: moving membranes to keep in shape［J］. Bioessays,2008,30(2):146-155.

［5］Blumenthal DK, Copps J, Smith-Nguyen EV, et al. The roles of the RIIbeta linker and N-terminal cyclic nucleotide-binding domain in determining the unique structures of the type IIbeta protein kinase A: a small angle x-ray and neutron scattering study［J］. J Biol Chem,2014,289(41):28505-28512.

［6］Colledge M, Scott JD. AKAPs: from structure to function［J］. Trends Cell Biol,1999,9(6):216-221.

［7］Cummings DE, Brandon EP, Planas JV, et al. Genetically lean mice result from targeted disruption of the RII beta subunit of protein kinase A［J］. Nature,1996,382(6592):622-626.［8］Ilouz R, Bubis J, Wu J, et al. Localization and quaternary structure of the PKA RIbeta holoenzyme［J］. Proc Natl Acad Sci USA,2012,109(31):12443-12448.

［9］Vigil D, Blumenthal DK, Taylor SS, et al. Solution scattering reveals large differences in the global structures of type Ⅱ protein kinase A isoforms［J］. J Mol Biol,2006,357(3):880-889.

［10］Kinderman FS, Kim C, von Daake S, et al. A dynamic mechanism for AKAP binding to RII isoforms of cAMP-dependent protein kinase［J］. Mol Cell,2006,24(3):397-408.

［11］Scott JD, Dessauer CW, Tasken K. Creating order from chaos: cellular regulation by kinase anchoring［J］. Annu Rev Pharmacol Toxicol,2013,53(1):187-210.

［12］Diskar M, Zenn HM, Kaupisch A, et al. Molecular basis for isoform-specific autoregulation of protein kinase A［J］. Cell Signal,2007,19(10):2024-2034.

［13］Martin BR, Deerinck TJ, Ellisman MH, et al. Isoform-specific PKA dynamics revealed by dye-triggered aggregation and DAKAP1alpha-mediated localization in living cells［J］. Chem Biol,2007,14(9):1031-1042.

［14］Canaves JM, Taylor SS. Classification and phylogenetic analysis of the cAMP-dependent protein kinase regulatory subunit family［J］. J Mol Evol,2002,54(1):17-29.

［15］Kim C, Cheng CY, Saldanha SA, et al. PKA-I holoenzyme structure reveals a mechanism for cAMP-dependent activation［J］. Cell,2007,130(6):1032-1043.

［16］Wu J, Brown SH, von Daake S, et al. PKA type IIalpha holoenzyme reveals a combinatorial strategy for isoform diversity［J］. Science,2007,318(5848):274-279.

［17］Herberg FW, Taylor SS, Dostmann WR. Active site mutations define the pathway for the cooperative activation of cAMP-dependent protein kinase［J］. Biochemistry,1996,35(9):2934-2942.

［18］Zhang P, Knape MJ, Ahuja LG, et al. Single turnover autophosphorylation cycle of the PKA RⅡbeta oloenzyme［J］. PLoS Biol,2015,13(7):e1002192.

［19］Oliveria SF, Dell’Acqua ML, Sather WA. AKAP79/150 anchoring of calcineurin controls neuronal L-type Ca2+ channel activity and nuclear signaling［J］. Neuron,2007,55(2):261-275.

［20］Rangel-Aldao R, Rosen OM. Dissociation and reassociation of the phosphorylated and nonphosphorylated forms of adenosine 3’:5’-monophosphate-dependent protein kinase from bovine cardiac muscle［J］. J Biol Chem,1976,251(11):3375-3380.

［21］Amieux PS, Cummings DE, Motamed K, et al. Compensatory regulation of RIalpha protein levels in protein kinase A mutant mice［J］. J Biol Chem,1997,272(7):3993-3998.

［22］Schreyer SA, Cummings DE, Mcknight GS, et al. Mutation of the RⅡbeta subunit of protein kinase A prevents diet-induced insulin resistance and dyslipidemia in mice［J］. Diabetes,2001,50(11):2555-2562.

［23］Enns LC, Morton JF, Treuting PR, et al. Disruption of protein kinase A in mice enhances healthy aging［J］. PLoS One,2009,4(6):e5963.

［24］Rosen ED, Macdougald OA. Adipocyte differentiation from the inside out［J］. Nat Rev Mol Cell Biol,2006,7(12):885-896.

［25］Farmer SR. Transcriptional control of adipocyte formation［J］. Cell Metab,2006,4(4):263-273.

［26］Peverelli E, Ermetici F, Corbetta S, et al. PKA regulatory subunit R2B is required for murine and human adipocyte differentiation［J］. Endocr Connect,2013,2(4):196-207.

［27］Cummings DE, Brandon EP, Planas JV, et al. Genetically lean mice result from targeted disruption of the RⅡ beta subunit of protein kinase A［J］. Nature,1996,382(6592):622-626.

［28］Meier MK, Alig L, BurgiSaville ME, et al. Phenethanolamine derivatives with calorigenic and antidiabetic qualities［J］. Int J Obes,1984,8 (Suppl 1):215-225.

［29］Dolan JA, Muenkel HA, Burns MG, et al. Beta-3 adrenoceptor selectivity of the dioxolane dicarboxylate phenethanolamines［J］. J Pharmacol Exp Ther,1994,269(3):1000-1006.

［30］Ek I, Arner P, Ryden M, et al. A unique defect in the regulation of visceral fat cell lipolysis in the polycystic ovary syndrome as an early link to insulin resistance［J］. Diabetes,2002,51(2):484-492.

［31］Newhall KJ, Cummings DE, Nolan MA, et al. Deletion of the RⅡbeta-subunit of protein kinase A decreases body weight and increases energy expenditure in the obese, leptin-deficient ob/ob mouse［J］. Mol Endocrinol,2005,19(4):982-991.

［32］Pelleymounter MA, Cullen MJ, Baker MB, et al. Effects of the obese gene product on body weight regulation in ob/ob mice［J］. Science,1995,269(5223):540-543.

［33］Halaas JL, Gajiwala KS, Maffei M, et al. Weight-reducing effects of the plasma protein encoded by the obese gene［J］. Science,1995,269(5223):543-546.

［34］Kirk EA, Moe GL, Caldwell MT, et al. Hyper-and hypo-responsiveness to dietary fat and cholesterol among inbred mice: searching for level and variability genes［J］. J Lipid Res,1995,36(7):1522-1532.

［35］Del Gbbo A, Peverelli E, Treppiedi D, et al. Expression of protein kinase A regulatory subunits in benign and malignant human thyroid tissues: a systematic review［J］. Exp Cell Res,2016,346(1):85-90.

［36］Almeida MQ, Stratakis CA. How does cAMP/protein kinase A signaling lead to tumors in the adrenal cortex and other tissues［J］? Mol Cell Endocrinol,2011,336(1-2):162-168.

［37］Mantovani G, Lania AG, Bondioni S, et al. Different expression of protein kinase A (PKA) regulatory subunits in cortisol-secreting adrenocortical tumors: relationship with cell proliferation［J］. Exp Cell Res,2008,314(1):123-130.

［38］Basso F, Rocchetti F, Rodriguez S, et al. Comparison of the effects of PRKAR1A and PRKAR2B depletion on signaling pathways, cell growth, and cell cycle control of adrenocortical cells［J］. Horm Metab Res,2014,46(12):883-888.

［39］Ferraro FR, Sparta DR, Knapp DJ, et al. Increased consumption but not operant self-administration of ethanol in mice lacking the RIIbeta subunit of protein kinase A［J］. Alcohol Clin Exp Res,2006,30(5):825-835.

［40］Thiele TE, Willis B, Stadler J, et al. High ethanol consumption and low sensitivity to ethanol-induced sedation in protein kinase A-mutant mice［J］. J Neurosci,2000,20(10):C75.

［41］Fee JR, Sparta DR, Knapp DJ, et al. Predictors of high ethanol consumption in RIIbeta knock-out mice: assessment of anxiety and ethanol-induced sedation［J］. Alcohol Clin Exp Res,2004,28(10):1459-1468.

［42］Belcher SM, Le HH, Spurling L, et al. Rapid estrogenic regulation of extracellular signal-regulated kinase 1/2 signaling in cerebellar granule cells involves a G protein-and protein kinase A-dependent mechanism and intracellular activation of protein phosphatase 2A［J］. Endocrinology,2005,146(12):5397-5406.

［43］Thiele TE, Sparta DR, Hayes DM, et al. A role for neuropeptide Y in neurobiological responses to ethanol and drugs of abuse［J］. Neuropeptides,2004,38(4):235-243.

［44］Shingo AS, Kito S. Estradiol induces PKA activation through the putative membrane receptor in the living hippocampal neuron［J］. J Neural Transm (Vienna),2005,112(11):1469-1473.

［45］Planas JV, Cummings DE, Idzerda RL, et al. Mutation of the RIIbeta subunit of protein kinase A differentially affects lipolysis but not gene induction in white adipose tissue［J］. J Biol Chem,1999,274(51):36281-36287.

[1]	贺继平苏晓云崔建梅张晓宇门杰周琛婓. 跑台运动对血管痴呆大鼠学习记忆的影响及其作用机制[J]. 解剖学报, 2023, 54(3): 276-282.
[2]	高赛红张小良杨迎春乔海兵. 高脂血症大鼠脑缺血/再灌注后p38 MAPK活化及对Bax和Bcl-2表达的影响[J]. 解剖学报, 2023, 54(1): 50-55.
[3]	王昱秦序何九军王逸璇 . 白肉灵芝水提物改善衰老大鼠认知功能的作用[J]. 解剖学报, 2022, 53(6): 711-718.
[4]	亓玉心杨文平刘爽韩蕃颉王海滨苏新云. 布地奈德对哮喘大鼠气道平滑肌细胞增殖及凋亡的影响[J]. 解剖学报, 2021, 52(5): 795-802.
[5]	金玲燕王弯弯张红陈琦. 不良心理应激对人卵巢癌荷瘤裸鼠皮下移植瘤生长及表皮生长因子受体、磷酸化蛋白激酶B、血管内皮生长因子蛋白表达的影响[J]. 解剖学报, 2018, 49(5): 617-623.
[6]	胡志红闫君宝李三强胡咏梅. 间歇性饥饿对亚硝酸钠所致大鼠海马神经细丝过度磷酸化及空间学习记忆损伤的改善作用[J]. 解剖学报, 2017, 48(1): 14-18.
[7]	汤諹李树清*. 缺血后适应调控树鼩脑缺血海马CA1区Akt（pS473）和Akt（pT308）过磷酸化的抗凋亡机制[J]. 解剖学报, 2016, 47(5): 591-598.
[8]	刘海莉朱德晓吴金涛岳庆伟王辉李泽岩李贵宝刘增训张静孙晋浩*. 细胞外信号调节激酶/miR-133b在甲基苯丙胺致PC12细胞神经毒性的作用及机制[J]. 解剖学报, 2014, 45(6): 755-760.
[9]	刘景利石新丽崔澂任来峰郑文广李明远*. p53-Ser³⁹²的磷酸化在肿瘤治疗中的作用[J]. 解剖学报, 2014, 45(3): 437-440.
[10]	刘文彬;惠珊珊;胡小慧;聂磊;马慧;王玲;肖亚梅;李万程;. 蛋白磷酸酶-2A催化亚基C在小鼠不同器官中的分化表达与定位[J]. , 2011, 42(3): 384-388.
[11]	朱蕙霞;金国华;田美玲;秦建兵;谭雪锋;金淑仪. 磷酸化细胞外信号调节激酶在海马提取液诱导神经干细胞向神经元分化过程中的表达[J]. , 2009, 40(6): 857-861.
[12]	邸菁;张凌志;柏树令. 磷酸化组蛋白H3——检测心肌细胞有丝分裂指数的可靠指标[J]. , 2008, 39(2): 272-274.
[13]	曾育琦;陈晓春;黄春;李永坤;彭小松;沈杰;黄天文. 人参皂苷Rg1通过调节GSK-3β/PP2A活性减轻皮层神经元Tau蛋白过度磷酸化[J]. , 2007, 38(6): 665-670.
[14]	郑关毅;陈晓春;刘昌云等. 一氧化氮激活应激活化蛋白激酶/c-Jun氨基末端激酶及p38MAP激酶诱导脑缺血再灌注后神经元凋亡[J]. , 2006, 37(6): 633-639.
[15]	刘能保;洪小平;李晓恒等. 慢性复合应激对学习记忆的影响及cAMP/PKA-CREB信号通路的作用[J]. , 2006, 37(6): 611-616.