Use of protein chip mass spectrometry to monitor biotinylation reactions.
Use of protein chip mass spectrometry to monitor biotinylation reactions.
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使用蛋白质芯片质谱法监测生物素化反应。
DOI：
10.1007/s00253-006-0710-1
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发表时间：
2007
影响因子：
5
通讯作者：
Boyle,MichaelDP
中科院分区：
文献类型：
--
作者：
Blazer,LeviL;Boyle,MichaelDP
表面增强激光解吸/电离飞行时间分析用于监测许多模型蛋白质和肽靶标生物素化过程中产物形成的动力学和异质性。所选分子是 IgG 结合蛋白、蛋白 A、人血清白蛋白和对应于链球菌 M1 蛋白 N 末端的合成肽。通过对天然材料的分子质量变化进行动力学分析来确定生物素化的程度。通过与 N-羟基琥珀酰亚胺生物素反应修饰的每个残基都会在天然蛋白质或多肽中添加~341 amu。该方法的新颖性在于能够确定分子量变化，而无需首先将目标分子与生物素化试剂分离。分析快速、简单，并提供了有关添加的生物素分子的平均数量和所得产品的均质性的信息。
Surface-enhanced laser desorption/ionization time-of-flight analysis was used to monitor both the kinetics and heterogeneity of product formation during the biotinylation of a number of model proteins and peptide targets. The selected molecules were the IgG-binding protein, protein A, human serum albumin, and a synthetic peptide corresponding to the N terminus of a streptococcal M1 protein. The extent of biotinylation was determined by kinetic analysis of the shift in molecular mass from the native material. Each residue modified by reaction withN-hydroxysuccinimide biotin resulted in an addition of ∼341 amu to the native protein or polypeptide. The novelty of the method was in the ability to determine the molecular mass shift, without first separating the targeted molecule from the biotinylating reagent. The analysis was rapid, simple, and provided information on the average number of biotin molecules added and the homogeneity of the resulting product.