The intracellular mobility of NPY and a putative mitochondrial form of NPY in neuronal cells.
Preproneuropeptide Y is a precursor peptide to mature neuropeptide Y (NPY), which is a universally expressed peptide in the central and peripheral nervous system. NPY is normally routed to endoplasmic reticulum and secretory vesicles in cells, which secrete NPY. In our previous studies, we found a functional Leucine7 to Proline7 (L7P) polymorphism in the signal peptide sequence of preproNPY. This polymorphism affects the secretion of NPY and causes multiple physiological effects in humans. The sequence of NPY mRNA contains two in frame kozak sequences that allow translation initiation to shift, and translation of two proteins. In addition to mature NPY(1-36) also a putative truncated NPY(17-36) with mitochondrial targeting signal is produced. The purpose of this study was to investigate the protein mobility of the putative mitochondrial fragment and the effect of the L7P polymorphism on the cellular level using GFP tagged constructs. The mobility was studied with fluorescence recovery after photobleaching technique in a neuronal cell line. We found that the mobility of the secretory vesicles with NPY(1-36) in cells with L7P genotype was increased in comparison to vesicle mobility in cells with the more abundant L7L genotype. The mobility in the cells with the putative mitochondrial construct was found to be very low. According to the results of the present study, the mitochondrial truncated peptide stays in the mitochondrion. It can be hypothesized that this could be one of the factors affecting energy balance of the membranes of the mitochondrion.
Kaipio
Katja
K
Department of Pharmacology, Drug Development and Therapeutics, University of Turku, PharmaCity, FIN-20520 Turku, Finland.
Pesonen
Ullamari
U
D045744:Cell Line, Tumor; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D007930:Leucine / Q000031:analogs & derivatives / Q000235:genetics; D018613:Microscopy, Confocal; D008928:Mitochondria / Q000378:metabolism*; D009447:Neuroblastoma / Q000473:pathology; D009474:Neurons / Q000378:metabolism / Q000648:ultrastructure*; D009478:Neuropeptide Y / Q000235:genetics / Q000378:metabolism*; D011110:Polymorphism, Genetic / Q000235:genetics; D011392:Proline / Q000235:genetics; D021381:Protein Transport / Q000235:genetics; D014162:Transfection / Q000379:methods
D009478:Neuropeptide Y; D049452:Green Fluorescent Proteins; C035766:leucinal; D011392:Proline; D007930:Leucine