Gentile Fabrizio, Pizzimenti Stefania, Arcaro Alessia, Pettazzoni Piergiorgio, Minelli Rosalba, D'Angelo Daniela, Mamone Gianfranco, Ferranti Pasquale, Toaldo Cristina, Cetrangolo Gianpaolo, Formisano Silvestro, Dianzani Mario U, Uchida Koji, Dianzani Chiara, Barrera Giuseppina
Dipartimento di Scienze per la Salute, Università del Molise, via De Sanctis, Campobasso 86100, Italy.
Biochem J. 2009 Aug 13;422(2):285-94. doi: 10.1042/BJ20090564.
HNE (4-hydroxynonenal), the major product of lipoperoxidation, easily reacts with proteins through adduct formation between its three main functional groups and lysyl, histidyl and cysteinyl residues of proteins. HNE is considered to be an ultimate mediator of toxic effects elicited by oxidative stress. It can be detected in several patho-physiological conditions, in which it affects cellular processes by addition to functional proteins. We demonstrated in the present study, by MS and confirmed by immunoblotting experiments, the formation of HNE-alpha-enolase adduct(s) in HL-60 human leukaemic cells. Alpha-enolase is a multifunctional protein that acts as a glycolytic enzyme, transcription factor [MBP-1 (c-myc binding protein-1)] and plasminogen receptor. HNE did not affect alpha-enolase enzymatic activity, expression or intracellular localization, and did not change the expression and localization of MBP-1 either. Confocal and electronic microscopy results confirmed the plasma membrane, cytosolic and nuclear localization of alpha-enolase in HL-60 cells and demonstrated that HNE was colocalized with alpha-enolase at the surface of cells early after its addition. HNE caused a dose- and time-dependent reduction of the binding of plasminogen to alpha-enolase. As a consequence, HNE reduced adhesion of HL-60 cells to HUVECs (human umbilical vein endothelial cells). These results could suggest a new role for HNE in the control of tumour growth and invasion.
4-羟基壬烯醛（HNE）是脂质过氧化的主要产物，它通过其三个主要功能基团与蛋白质的赖氨酰、组氨酰和半胱氨酰残基之间形成加合物，从而很容易与蛋白质发生反应。HNE被认为是氧化应激引发的毒性作用的最终介质。在几种病理生理条件下都能检测到它，在这些条件下，它通过与功能性蛋白质结合来影响细胞过程。在本研究中，我们通过质谱法进行了证明，并通过免疫印迹实验予以证实，在HL-60人白血病细胞中形成了HNE-α-烯醇化酶加合物。α-烯醇化酶是一种多功能蛋白质，它作为糖酵解酶、转录因子[MBP-1（c-myc结合蛋白-1）]和纤溶酶原受体发挥作用。HNE不影响α-烯醇化酶的酶活性、表达或细胞内定位，也不改变MBP-1的表达和定位。共聚焦显微镜和电子显微镜结果证实了α-烯醇化酶在HL-60细胞中的质膜、胞质和核定位，并表明在添加HNE后早期，HNE与α-烯醇化酶在细胞表面共定位。HNE导致纤溶酶原与α-烯醇化酶结合呈剂量和时间依赖性减少。因此，HNE降低了HL-60细胞与人类脐静脉内皮细胞（HUVECs）的黏附。这些结果可能提示HNE在肿瘤生长和侵袭控制中具有新的作用。
