The Role of Intestinal Fatty Acid Binding Proteins in Protecting Cells from Fatty Acid Induced Impairment of Mitochondrial Dynamics and Apoptosis

Abstract

Background/Aims: The conformation, folding and lipid binding properties of the intestinal fatty acid binding proteins (IFABP) have been extensively investigated. In contrast, the functional aspects of these proteins are not understood and matter of debates. In this study, we aim to address the deleterious effects of FA overload on cellular components, particularly mitochondria; and how IFABP helps in combating this stress by restoring the mitochondrial dynamics. Methods: In the present study the functional aspect of IFABP under conditions of lipid stress was studied by a string of extensive in-cell studies; flow cytometry by fluorescence-activated cell sorting (FACS), confocal imaging, western blotting and quantitative real time PCR. We deployed ectopic expression of IFABP in rescuing cells under the condition of lipid stress. Again in order to unveil the mechanistic insights of functional traits, we arrayed extensive computational approaches by means of studying centrality calculations along with protein-protein association and ligand induced cluster dissociation. While addressing its functional importance, we used FCS and in-silico computational analyses, to show the structural distribution and the underlying mechanism of IFABP’s action. Results: Ectopic expression of IFABP in HeLa cells has been found to rescue mitochondrial morphological dynamics and restore membrane potential, partially preventing apoptotic damage induced by the increased FAs. These findings have been further validated in the functionally relevant intestinal Caco-2 cells, where the native expression of IFABP protects mitochondrial morphology from abrogation induced by FA overload. However, this native level expression is insufficient to protect against apoptotic cell death, which is rescued, at least partially in cells overexpressing IFABP. In addition, shRNA mediated IFABP knockdown in Caco-2 cells compromises mitochondrial dynamics and switches on intrinsic apoptotic pathways under FA-induced metabolic stress. Conclusion: To summarize, the present study implicates functional significance of IFABP in controlling ligand-induced damage in mitochondrial dynamics and apoptosis.

Development and in vitro characterization of capecitabine loaded biopolymeric vehicle for the treatment of colon cancer

“…HT29 Human colon cancer cell line was purchased from the National Centre for Cell Sciences (NCCS), Pune, India and were maintained in Dulbecco's Modified Eagle's Medium (DMEM), which was supplemented with 10% heat inactivated fetal bovine serum (FBS), 4.5 g/L glucose, 4 mM glutamine, 110 mg/L sodium pyruvate, 1.5 g/L sodium bicarbonate, 50 μg/ml streptomycin and 50 μg/ml penicillin 35,36 . HT29 cells were incubated in the humidified air containing 5% of CO 2 at 37°C.…”

“…fetal bovine serum (FBS), 4.5 g/L glucose, 4 mM glutamine, 110 mg/L sodium pyruvate, 1.5 g/L sodium bicarbonate, 50 μg/ml streptomycin and 50 μg/ml penicillin 35,36. HT29 cells were incubated in the humidified air containing 5% of CO 2 at 37 C. Cells were replenished and sub-cultured as per ATCC recommendations (ATCC, Manassus, VA).37,38 2.2.15 | Cell viability assay…”

The Interplay of Iron and Lipid Homeostasis in Non-Alcoholic Fatty Liver Disease

“…FABP1 is the most abundant FABP in hepatocytes and represents up to 5% of all cytosolic proteins (75,76). FFAs are known as cytotoxic to cells (77,78), and their binding to FABPs aids in decreasing their toxicity (79). Additionally, changes in FABP1 expression substantially affect the rate of fatty acid uptake, a property that other FABPs do not demonstrate (80,81).…”

An exploration of the SARS-CoV-2 spike receptor binding domain (RBD) – a complex palette of evolutionary and structural features

“…In order to determine the structure of the ACE2-RBD complex and to identify the residue/segments of the RBD interacting with the ACE2 receptor protein, we resorted the computational strategy of protein-protein docking (30). In this approach, the coordinate information of the ACE2 receptor protein was resorted from protein data bank (PDB) (PDB ID: 1R42).…”