Fujimoto Shiori, Murakami Yoto, Miyake Haruna, Hayase Fumitaka, Watanabe Hirohito
a Department of Agriculture , Meiji University , Kawasaki , Japan.
Biosci Biotechnol Biochem. 2019 Jun;83(6):1136-1145. doi: 10.1080/09168451.2019.1585745. Epub 2019 Mar 1.
Advanced glycation end products (AGEs) are implicated in the development of diabetic complications via the receptor for AGEs (RAGE). We have reported that the 3-hydroxypyridinium (3HP)-containing AGEs derived from α-hydroxyaldehydes physically interact with RAGE and show cytotoxicity. Lactaldehyde (LA) is formed from a reaction between threonine and myeloperoxidase, but no LA-derived AGEs have been characterized. Here, we identify the structure and physiological effects of an AGE derived from LA. We isolated a novel 3HP derivative, 2-acetamido-6-(3-hydroxy-5-methyl-pyridin-1-ium-1-yl)hexanoate, named as N-acetyl-LAPL (lactaldehyde-derived pyridinium-type lysine adduct), from a mixture of LA with N-acetyl-L-lysine. LAPL was also detected in the LA-modified protein. LAPL elicited toxicity in PC12 neuronal cells, but the effect was suppressed by the soluble form of RAGE as a decoy receptor. Moreover, surface plasmon resonance-based analysis revealed that LAPL specifically binds to recombinant RAGE. These results indicate that LA generates an AGE containing the 3HP moiety and contributes to RAGE-dependent cytotoxicity. Abbreviations: AGEs: advanced glycation end products; RAGE: receptor for advanced glycation end products; 3HP: 3-hydroxypyridinium; LA: lactaldehyde; LAPL: lactaldehyde-derived pyridinium-type lysine adduct; BSA: bovine serum albumin; GLAP: glyceraldehyde-derived pyridinium; MPO: myeloperoxidase; HFBA: heptafluorobutyric acid; TFA: trifluoroacetic acid; HPLC: high performance liquid chromatography; LC-ESI-QTOF-MS: liquid chromatography-electrospray ionization-quadrupole time-of-flight-mass spectrometry; NMR: nuclear magnetic resonance; LA-BSA: lactaldehyde-modified bovine serum albumin; PBS: phosphate buffered saline, GST, glutathione S-transferase; SPR: surface plasmon resonance; OP-lysine: 2-ammonio-6-(3-oxidopyridinium-1-yl)hexanoate; GLO1: glyoxalase 1; MG, methylglyoxal.
晚期糖基化终末产物（AGEs）通过晚期糖基化终末产物受体（RAGE）参与糖尿病并发症的发生发展。我们曾报道，源自α-羟基醛的含3-羟基吡啶鎓（3HP）的AGEs与RAGE发生物理相互作用并表现出细胞毒性。乳醛（LA）由苏氨酸与髓过氧化物酶反应形成，但尚未对源自LA的AGEs进行表征。在此，我们鉴定了源自LA的一种AGE的结构和生理效应。我们从LA与N-乙酰-L-赖氨酸的混合物中分离出一种新型3HP衍生物，2-乙酰氨基-6-（3-羟基-5-甲基吡啶-1-鎓-1-基）己酸，命名为N-乙酰-LAPL（乳醛衍生的吡啶型赖氨酸加合物）。在LA修饰的蛋白质中也检测到了LAPL。LAPL在PC12神经元细胞中引发毒性，但该效应被作为诱饵受体的可溶性RAGE所抑制。此外，基于表面等离子体共振的分析表明LAPL特异性结合重组RAGE。这些结果表明LA产生了一种含3HP部分的AGE，并导致了RAGE依赖性细胞毒性。缩写：AGEs：晚期糖基化终末产物；RAGE：晚期糖基化终末产物受体；3HP：3-羟基吡啶鎓；LA：乳醛；LAPL：乳醛衍生的吡啶型赖氨酸加合物；BSA：牛血清白蛋白；GLAP：甘油醛衍生的吡啶鎓；MPO：髓过氧化物酶；HFBA：七氟丁酸；TFA：三氟乙酸；HPLC：高效液相色谱；LC-ESI-QTOF-MS：液相色谱-电喷雾电离-四极杆飞行时间质谱；NMR：核磁共振；LA-BSA：乳醛修饰的牛血清白蛋白；PBS：磷酸盐缓冲盐水；GST：谷胱甘肽S-转移酶；SPR：表面等离子体共振；OP-赖氨酸：2-氨-6-（3-氧化吡啶鎓-1-基）己酸；GLO1：乙二醛酶1；MG：甲基乙二醛。
