title: Overexpression of myosin-IIB in the brain of a rat model of streptozotocin-induced diabetes.
abstract: The Ca(2+)/calmodulin complex interacts with and regulates various enzymes and target proteins known as calmodulin-binding proteins (CaMBPs). This group of proteins includes molecular motors such as myosins. In this study, we show that non-muscle myosin-IIB is overexpressed in the brains of diabetic rats. We isolated CaMBPs from the brains of non-diabetic rats and rats with streptozotocin-induced diabetes and purified them by immobilized-calmodulin affinity chromatography. The proteins were eluted with EGTA and urea, separated by SDS-PAGE, digested and submitted to peptide mass fingerprinting analysis. Thirteen intense bands were found in both types of brains, two were found exclusively in non-diabetic brains and four were found exclusively in diabetic brains. A large fraction of the eluted proteins contained putative IQ motifs or calmodulin-binding sites. The results of the myosin-IIB affinity chromatography elution, western blot and RT-PCR analyses suggest that myosin-IIB protein and mRNA are expressed at high levels in diabetic brains. This is the first study that has demonstrated differential expression of CaMBPs in diabetic and non-diabetic brain tissue through a comparative proteomic analysis, and it opens up a new approach to studying the relationship between the expression of myosins in the brain, hyperglycemia and intracellular calcium regulation.
authors.affiliation: Institute of Genetics and Biochemistry, Federal University of Uberlândia, Campus Umuarama, 38400-902, Uberlândia-MG, Brazil.
authors.forename: Luciana Karen
authors.initials: LK
authors.lastname: Calábria
authors.forename: Gabriel Costa Nunes
authors.initials: GC
authors.lastname: da Cruz
authors.forename: Rafael
authors.initials: R
authors.lastname: Nascimento
authors.forename: Washington João
authors.initials: WJ
authors.lastname: Carvalho
authors.forename: Neire Moura
authors.initials: NM
authors.lastname: de Gouveia
authors.forename: Fernanda Vieira
authors.initials: FV
authors.lastname: Alves
authors.forename: Fabiana Barcelos
authors.initials: FB
authors.lastname: Furtado
authors.forename: Hellen Cristina
authors.initials: HC
authors.lastname: Ishikawa-Ankerhold
authors.forename: Marcelo Valle
authors.initials: MV
authors.lastname: de Sousa
authors.forename: Luiz Ricardo
authors.initials: LR
authors.lastname: Goulart
authors.forename: Foued Salmen
authors.initials: FS
authors.lastname: Espindola
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D001923:Brain Chemistry / Q000502:physiology*; D002148:Calmodulin-Binding Proteins / Q000378:metabolism; D002846:Chromatography, Affinity; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D003921:Diabetes Mellitus, Experimental / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D007150:Immunohistochemistry; D008297:Male; D008969:Molecular Sequence Data; D024501:Nonmuscle Myosin Type IIB / Q000096:biosynthesis*; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry; D012313:RNA / Q000096:biosynthesis / Q000302:isolation & purification; D051381:Rats; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014357:Trypsin / Q000737:chemistry
chemical_list: D002148:Calmodulin-Binding Proteins; D018076:DNA, Complementary; D019151:Peptide Library; D010455:Peptides; D012313:RNA; D014357:Trypsin; D024501:Nonmuscle Myosin Type IIB