1. Overexpression of myosin-IIB in the brain of a rat model of streptozotocin-induced diabetes
- Author
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Gabriel Costa Nunes da Cruz, Neire Moura de Gouveia, Luciana Karen Calábria, Washington João de Carvalho, Foued Salmen Espindola, Hellen Ishikawa-Ankerhold, Luiz Ricardo Goulart, Marcelo Valle de Sousa, Fernanda Vieira Alves, Fabiana Barcelos Furtado, and Rafael Nascimento
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Male ,DNA, Complementary ,Calmodulin ,Blotting, Western ,Molecular Sequence Data ,Clinical Neurology ,Chromatography, Affinity ,Diabetes Mellitus, Experimental ,chemistry.chemical_compound ,Diabetes mellitus ,Western blot ,Peptide mass fingerprinting ,Affinity chromatography ,Peptide Library ,Myosin ,medicine ,Animals ,Trypsin ,Amino Acid Sequence ,Rats, Wistar ,Brain Chemistry ,Messenger RNA ,Nonmuscle Myosin Type IIB ,medicine.diagnostic_test ,biology ,Reverse Transcriptase Polymerase Chain Reaction ,Brain ,Streptozotocin ,Immunohistochemistry ,Rats ,EGTA ,Neurology ,chemistry ,Biochemistry ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Hyperglycemia ,biology.protein ,RNA ,Calmodulin-Binding Proteins ,Electrophoresis, Polyacrylamide Gel ,Calcium ,Myosin-IIB ,Neurology (clinical) ,Peptides ,medicine.drug - 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.
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