Robinson N E, Robinson A B
Department of Chemistry, California Institute of Technology, Pasadena, CA 91125, USA.
Proc Natl Acad Sci U S A. 2001 Jan 30;98(3):944-9. doi: 10.1073/pnas.98.3.944.
A convenient and precise mass spectrometric method for measurement of the deamidation rates of glutaminyl and asparaginyl residues in peptides and proteins has been developed; the rates of deamidation of 306 asparaginyl sequences in model peptides at pH 7.4, 37.0 degrees C, 0.15 M Tris.HCl buffer have been determined; a library of 913 amide-containing peptides for use by other investigators in similar studies has been established; and, by means of simultaneous deamidation rate measurements of rabbit muscle aldolase and appropriate model peptides in the same solutions, the use of this method for quantitative measurement of the relative effects of primary, secondary, tertiary, and quaternary protein structure on deamidation rates has been demonstrated. The measured rates are discussed with respect to the hypothesis that glutaminyl and asparaginyl residues serve, through deamidation, as molecular timers of biological events.
已开发出一种便捷且精确的质谱方法,用于测量肽和蛋白质中谷氨酰胺基和天冬酰胺基残基的脱酰胺速率;已测定了在pH 7.4、37.0℃、0.15 M Tris.HCl缓冲液中模型肽中306个天冬酰胺基序列的脱酰胺速率;已建立了一个由913个含酰胺肽组成的文库,供其他研究人员用于类似研究;并且,通过在相同溶液中同时测量兔肌肉醛缩酶和合适模型肽的脱酰胺速率,已证明该方法可用于定量测量蛋白质一级、二级、三级和四级结构对脱酰胺速率的相对影响。针对谷氨酰胺基和天冬酰胺基残基通过脱酰胺作用作为生物事件分子定时器的假说,对所测得的速率进行了讨论。