Microheterogeneity of the major grass group 6 allergen Phl p 6: Analysis by mass spectrometry
Microheterogeneity of the major grass group 6 allergen Phl p 6: Analysis by mass spectrometry
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DOI：
10.1002/pmic.200300706
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发表时间：
2004-05-01
期刊：
影响因子：
3.4
通讯作者：
Petersen, A
中科院分区：
文献类型：
--
作者：
Blume, C;Lindner, B;Petersen, A
过敏原的精确结构表征是改善诊断和找到针对过敏性疾病的治疗策略的基本要求。天然草花粉过敏原表现出各种各样的亚型，它仍然是未知的，是否这种微观异质性是必不可少的过敏反应或致敏功能的影响。明确定义的重组过敏原被认为是替代天然过敏原的临床试验。对于主要的梯牧草花粉过敏原Phl p6通过双向电泳（2-DE）、高分辨电喷雾离子化-傅立叶变换-质谱（ESI-MS）和免疫印迹（ELISA）等方法，（ESI-FT-MS）的完整分子，以及通过使用基质辅助激光解吸/电离飞行时间质谱法（MALDI-TOF-MS）的胰蛋白酶肽质量指纹图谱。天然Phl p 6主要是两种异构体的混合物，这两种异构体相差两个氨基酸，导致5 Da的质量差异。对于这两种异构体中的每一种，鉴定出在C-和/或N-末端具有修饰的六种变体。重组Phl p 6包含与主要同种型之一相同的结构，表明其代表天然Phl p 6的主要部分。
The precise structural characterization of allergens is a basic requirement to improve diagnostics and to find therapeutic strategies against allergic disorders. Natural grass pollen allergens exhibit a wide variety of isoforms and it is still unknown whether this micro heterogeneity is essential for the allergic reaction or has a functional effect on sensitization. Well-defined recombinant allergens are considered to replace natural allergens for clinical trials. For the major timothy grass pollen allergen Phl p 6 (similar to12 kDa) and a recombinant rPhl p 6 we determined the structural microheterogeneity by two-dimensional electrophoresis (2-DE), high-resolution electrospray ionization-Fourier transform-mass spectrometry (ESI-FT-MS) of the intact molecules, and by tryptic peptide mass fingerprinting using matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS). Natural Phl p 6 is a mixture of mainly two isoforms that differ by two amino acids leading to a mass difference of 5 Da. For each of this two isoforms six variants were identified with modifications at the C- and/or N-terminus. The recombinant Phl p 6 comprises the same structure as one of the main isoforms indicating that it represents a major part of the natural Phl p 6.