Bachmann M, Huber J L, Liao P C, Gage D A, Huber S C
U.S. Department of Agriculture, Agriculture Research Service, Raleigh, NC 27695-7631, USA.
FEBS Lett. 1996 Jun 3;387(2-3):127-31. doi: 10.1016/0014-5793(96)00478-4.
The inhibitor protein (IP) that inactivates spinach leaf NADH:nitrate reductase (NR) has been identified for the first time as a member of the eukaryotic 14-3-3 protein family based on three lines of evidence. First, the sequence of an eight amino acid tryptic peptide, obtained from immunopurified IP, matched that of a highly conserved region of the 14-3-3 proteins. Second, an authentic member of the 14-3-3 family, recombinant Arabidopsis GF14omega, caused inactivation of phospho-NR in a magnesium-dependent manner identical to IP. Third, an anti-GF14 monoclonal antibody cross-reacted with IP and anti-IP monoclonal antibodies cross-reacted with GF14omega.
基于三条证据，首次确定使菠菜叶NADH：硝酸还原酶（NR）失活的抑制蛋白（IP）是真核生物14-3-3蛋白家族的成员。第一，从免疫纯化的IP获得的一个八氨基酸胰蛋白酶肽段的序列，与14-3-3蛋白高度保守区域的序列相匹配。第二，14-3-3家族的一个正宗成员，重组拟南芥GF14ω，以与IP相同的镁依赖方式导致磷酸化NR失活。第三，一种抗GF14单克隆抗体与IP发生交叉反应，抗IP单克隆抗体与GF14ω发生交叉反应。
