Glucose autoxidation induces functional damage to proteins via modification of critical arginine residues.
Glucose autoxidation induces functional damage to proteins via modification of critical arginine residues.
复制标题
DOI：
10.1021/bi200757d
复制
发表时间：
2011-07-12
期刊：
影响因子：
2.9
通讯作者：
Voziyan, Paul
中科院分区：
文献类型：
--
作者：
Chetyrkin, Sergei;Mathis, Missy;Pedchenko, Vadim;Sanchez, Otto A.;McDonald, W. Hayes;Hachey, David L.;Madu, Hartman;Stec, Donald;Hudson, Billy;Voziyan, Paul
高血糖蛋白的非酶修饰是引起糖尿病并发症的主要机制。当这些修饰靶向活性位点残基，从而影响蛋白质功能时，可产生致病性后果。在本研究中，我们以溶菌酶和胶原IV的RGD-α3NC1结构域为模型蛋白，在体外研究了葡萄糖自氧化在功能性蛋白损伤中的作用。我们证明，葡萄糖自氧化通过活性羰基物质（RCS）乙二醛（GO）和甲基乙二醛修饰关键精氨酸残基，诱导溶菌酶活性的抑制以及NC1结构域与αVβ3整合素受体的结合，而非氧化葡萄糖对蛋白质的内聚不影响蛋白质的功能。使用pyridoxamine阻断葡萄糖自氧化和RCS的产生，从而保护蛋白质功能，即使在高浓度葡萄糖存在的情况下，也证实了RCS在蛋白质损伤中的作用。葡萄糖自氧化可能导致体内蛋白质损伤，因为在糖尿病大鼠肾ECM分离的胶原IV NC1结构域的组装界面中检测到go衍生的精氨酸残基修饰水平增加。由于精氨酸残基经常存在于蛋白质活性位点，葡萄糖自氧化可能是高血糖和氧化环境下导致ECM蛋白功能损伤的共同机制。我们的数据还指出了功能研究的缺陷，特别是在细胞培养实验中，涉及葡萄糖处理，但没有考虑到葡萄糖自氧化引起的RCS的毒性作用。
Non-enzymatic modification of proteins in hyperglycemia is a major mechanism causing diabetic complications. These modifications can have pathogenic consequences when they target active site residues, thus affecting protein function. In the present study, we examined the role of glucose autoxidation in functional protein damage using lysozyme and RGD-α3NC1 domain of collagen IV as model proteins in vitro. We demonstrated that glucose autoxidation induced inhibition of lysozyme activity as well as NC1 domain binding to αVβ3 integrin receptor via modification of critical arginine residues by reactive carbonyl species (RCS) glyoxal (GO) and methylglyoxal while non-oxidative glucose adduction to the protein did not affect protein function. The role of RCS in protein damage was confirmed using pyridoxamine which blocked glucose autoxidation and RCS production, thus protecting protein function, even in the presence of high concentrations of glucose. Glucose autoxidation may cause protein damage in vivo since increased levels of GO-derived modifications of arginine residues were detected within the assembly interface of collagen IV NC1 domains isolated from renal ECM of diabetic rats. Since arginine residues are frequently present within protein active sites, glucose autoxidation may be a common mechanism contributing to ECM protein functional damage in hyperglycemia and oxidative environment. Our data also point out the pitfalls in functional studies, particularly in cell culture experiments, that involve glucose treatment but do not take into account toxic effects of RCS derived from glucose autoxidation.
登录
查看更多内容
影响因子：
3.5
作者：
Halliwell, B
通讯作者：
Halliwell, B
影响因子：
7.8
作者：
GEHLSEN, KR;SRIRAMARAO, P;SKUBITZ, APN
通讯作者：
SKUBITZ, APN
影响因子：
1.6
作者：
Kawamura, Shunsuke;Chijiiwa, Yuki;Torikata, Takao
通讯作者：
Torikata, Takao
影响因子：
2.9
作者：
GRAF, J;OGLE, RC;KLEINMAN, HK
通讯作者：
KLEINMAN, HK
影响因子：
4.1
作者：
Ahmed, MU;Frye, EB;Baynes, JW
通讯作者：
Baynes, JW