Behrens Maik, Margolis Joyce W, Margolis Frank L
Department of Anatomy and Neurobiology, University of Maryland School of Medicine, 685 West Baltimore Street, Baltimore, MD 21201, USA.
J Neurochem. 2003 Sep;86(5):1289-96. doi: 10.1046/j.1471-4159.2003.01940.x.
Olfactory marker protein (OMP) expression is a hallmark of mature vertebrate olfactory receptor neurons (ORNs). Evidence for OMP function derives from altered behavioral and electrophysiological activities of OMP-KO mice. The molecular basis for the altered phenotype following the deletion of OMP is still unclear. Recent structural studies predict the involvement of OMP in protein-protein interaction. Here we report the identification of an OMP partner, Bex2, by phage-display screening of an olfactory mucosal cDNA-library. In situ hybridization demonstrates cellular co-localization of OMP mRNA with mRNAs for Bex1, Bex2, and Bex3 in ORNs of olfactory tissue of the mouse. The OMP/Bex interaction has been confirmed by demonstrating the chemical cross-linking of recombinant rat OMP with a synthetic peptide derived from the Bex amino acid sequence. The subcellular localization of Bex and OMP proteins was evaluated in transfected HEK293 cells. Bex is visualized in the nucleus and cytoplasm. Following co-transfection we observed the unexpected presence of some OMP in the nucleus along with Bex. Together, these data argue convincingly that we have identified Bex as an OMP partner whose further characterization will provide insight to the role of OMP and to the mechanism of the OMP/Bex interaction in ORN differentiation and function.
嗅觉标记蛋白(OMP)的表达是成熟脊椎动物嗅觉受体神经元(ORN)的一个标志。OMP功能的证据来源于OMP基因敲除小鼠行为和电生理活动的改变。OMP缺失后表型改变的分子基础仍不清楚。最近的结构研究预测OMP参与蛋白质-蛋白质相互作用。在此,我们报告通过对嗅觉黏膜cDNA文库进行噬菌体展示筛选,鉴定出OMP的一个伙伴蛋白Bex2。原位杂交显示在小鼠嗅觉组织的ORN中,OMP mRNA与Bex1、Bex2和Bex3的mRNA在细胞中共定位。通过证明重组大鼠OMP与源自Bex氨基酸序列的合成肽发生化学交联,证实了OMP与Bex的相互作用。在转染的HEK293细胞中评估了Bex和OMP蛋白的亚细胞定位。Bex在细胞核和细胞质中可见。共转染后,我们观察到一些OMP与Bex一起意外地出现在细胞核中。总之,这些数据令人信服地表明,我们已将Bex鉴定为OMP的伙伴蛋白,对其进一步的表征将为OMP的作用以及OMP/Bex相互作用在ORN分化和功能中的机制提供深入了解。