Record Details

MOLECULAR CHARACTERIZATION AND IN SILICO ANALYSIS OF BUFFALO KLRF1 GENE

KrishiKosh

View Archive Info
 
 
Field Value
 
Title MOLECULAR CHARACTERIZATION AND IN SILICO ANALYSIS OF BUFFALO KLRF1 GENE
 
Creator Sendha, Jasobanta
 
Contributor Balabantaray, S
 
Subject KLRF1; KLRF2; CLEC2B; CLEC2A; Natural killer Cells; KLRF1-CLEC2B Interaction; KLRF2-CLEC2A Interaction; Molecular Dynamics Simulation
 
Description Killer cell lectin-like receptor subfamily F member 1 (KLRF1) or, NKp80 is an cysteine-linked homodimeric C-type lectin-like receptor (CTLR), is expressed in natural killer (NK) cells and stimulates cellular cytotoxicity. This receptor is encoded in Natural Killer Gene Complex (NKC) adjacent to its ligand CLEC2B (AICL) in a tail to tail orientation. It has been studied that the interaction mediated by KLRF1 and CLEC2B not only induces cellular cytotoxicity but also is crucially involved in activating the NK cell-monocyte crosstalk and stimulating the proinflammatory cytokines, IFNγ and TNF. This receptor is absent in rodent species and conserved in human. The detailed molecular mechanisms behind KLRF1 gene was studied in human; however no such studies has been done in ruminant species, such as cattle or, buffaloes. Here, to understand its genomic organization and its interaction with AICL, the full length buffalo KLRF1 mRNA was amplified from whole blood, cloned and sequenced. The genomic organization was inferred from UCSC genome browser search against cow genome and found in chromosome 4 and consists of 6 exons. The phylogenetic analysis with available orthologous revealed its presence in ruminant
6
cluster. Moreover, to understand the interaction of KLRF1 with CLEC2B, the structural bioinformatics approach was followed that includes, molecular modeling, docking and molecular dynamics simulation. First, the 3D models of CTLR domain of buKLRF1-CTLR and buCLEC2B (retrieved from NCBI protein database) was predicted using MODELLER and the built models were validated by analyzing their stereochemical parameters and model quality. The structural properties of the built models were also studied by performing 10ns MD simulation. Later, to understand the interaction mediated by buKLRF1 and CLEC2B, molecular docking was performed in reference to KLRF2-CLEC2A crystal structure. Now, to check the stability of docked complex, 10ns MD simulation was performed and the result was compared with KLRF2-CLEC2A. Analysis of backbone stability, compactness and total numbers H-bonds formed by the buKLRF1-CLEC2B indicated a complex stability. Again it was observed that the mode of interaction mediated by buKLRF1-CLEC2B found to very similar to the experimental structure of human KLRF2-CLEC2A. Overall, this study gives an idea to understand the presence of KLRF1 in buffalo and will helpful for future cell cytotoxicity studies in ruminant species.
 
Date 2017-01-03T14:31:55Z
2017-01-03T14:31:55Z
2016
 
Type Thesis
 
Identifier http://krishikosh.egranth.ac.in/handle/1/94163
 
Language en
 
Relation Th;4640
 
Format application/pdf