Dufresne C P, Wood T D, Hendrickson C L
Department of Chemistry, State University of New York at Buffalo 14260-3000, USA.
J Am Soc Mass Spectrom. 1998 Nov;9(11):1222-5. doi: 10.1016/S1044-0305(98)00102-0.
Glucokinase (GK, EC 2.7.1.2), a member of the enzyme family of hexokinases, has been shown to be linked to maturity-onset diabetes of the young type II (MODY-2). Although nucleotide and amino acid sequence information are available for the human varieties, they are not known for the variety from Bacillus stearothermophilus, which is often used in protein binding studies. Here, a combination of electrospray Fourier transform mass spectrometry (FTMS) and infrared multiphoton dissociation (IRMPD) is used to obtain accurate molecular weight and preliminary amino acid sequence information for the protein. Electrospray FTMS provides evidence of a solution phase dimer. In addition, dithiothreitol reduction shows no shift in high-resolution isotopic distributions, indicating a probable absence of disulfide bonds in the protein. The partial sequence information obtained from IRMPD could be the basis for creating a DNA probe for the protein.
葡萄糖激酶（GK，EC 2.7.1.2）是己糖激酶酶家族的成员，已被证明与青年型II型成年发病型糖尿病（MODY - 2）有关。虽然已有人类各种葡萄糖激酶的核苷酸和氨基酸序列信息，但常用于蛋白质结合研究的嗜热栖热芽孢杆菌的葡萄糖激酶的此类信息尚不清楚。在此，采用电喷雾傅里叶变换质谱（FTMS）和红外多光子解离（IRMPD）相结合的方法来获取该蛋白质的精确分子量和初步氨基酸序列信息。电喷雾FTMS提供了溶液相二聚体的证据。此外，二硫苏糖醇还原显示高分辨率同位素分布没有变化，表明该蛋白质中可能不存在二硫键。从IRMPD获得的部分序列信息可能是创建该蛋白质DNA探针的基础。
