Optimized proteomic analysis of a mouse model of cerebellar dysfunction using amine-specific isobaric tags
| Autor: | Kenneth M. Wannemacher, Jin Qian, Stella Elkabes, Yan Li, Christa M. Patterson, Michael P. Kurnellas, Sanqiang Pan, Oleg Borisov, Longwen Deng, Tong Liu, Hong Li, Jun Hu |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2006 |
| Předmět: |
Proteomics
Cerebellum Calbindins Ataxia Blotting Western Calcium-Transporting ATPases Biology Bioinformatics Biochemistry Calbindin Article Mice Plasma Membrane Calcium-Transporting ATPases S100 Calcium Binding Protein G Cerebellar Diseases medicine Animals Electrophoresis Gel Two-Dimensional Amines Shotgun proteomics Molecular Biology Cation Transport Proteins Mice Knockout Neurodegeneration Computational Biology Proteins Reproducibility of Results medicine.disease Chromatography Ion Exchange Transport protein Cell biology Mice Inbred C57BL Disease Models Animal medicine.anatomical_structure Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Plasma membrane Ca2+ ATPase sense organs medicine.symptom |
| Popis: | Recent proteomic applications have demonstrated their potential for revealing the molecular mechanisms underlying neurodegeneration. The present study quantifies cerebellar protein changes in mice that are deficient in plasma membrane calcium ATPase 2 (PMCA2), an essential neuronal pump that extrudes calcium from cells and is abundantly expressed in Purkinje neurons. PMCA2null mice display motor dyscoordination and unsteady gait deficits observed in neurological diseases such as multiple sclerosis and ataxia. We optimized an amine-specific isobaric tags (iTRAQ ™ )-based shotgun proteomics workflow for this study. This workflow took consideration of analytical variance as a function of ion signal intensity and employed biological repeats to aid noise reduction. Even with stringent protein identification criteria, we could reliably quantify nearly 1000 proteins, including many neuronal proteins that are important for synaptic function. We identified 21 proteins that were differentially expressed in PMCA2-null mice. These proteins are involved in calcium homeostasis, cell structure and chromosome organization. Our findings shed light on the molecular changes that underlie the neurological deficits observed in PMCA2-null mice. The optimized workflow presented here will be valuable for others who plan to implement the iTRAQ method. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|