| چکیده انگلیسی مقاله |
Background: When Staphylococcus aureus is grown in the presence of high concentration of external glucose, this sugar is phosphorylated by glucokinase (glkA) to form glucose-6-phosphate. This product subsequently enters into anabolic phase, which favors biofilm formation. The presence of ROK (repressor protein, open reading frame, sugar kinase) motif, phosphate-1 and -2 sites, and tyrosine kinase sites in glkA of S. aureus indicates that phosphorylation must regulate the glkA activity. The aim of the present study was to identify the effect of phosphorylation on the function of S. aureus glkA and biofilm formation. Methods: Pure glkA and protein-tyrosine kinase (BYK) of S. aureus ATCC 12600 were obtained by fractionating the cytosolic fractions of glkA1 and BYK-1 expressing recombinant clones through nickel metal chelate column. The pure glkA was used as a substrate for BYK and the phosphorylation of glkA was confirmed by treating with reagent A and resolving in SDS-PAGE, as well as staining with reagent A. The kinetic parameters of glkA and phosphorylated glkA were determined spectrophotometrically, and in silico tools were used for validation. S. aureus was grown in brain heart infusion broth, which was supplemented with glucose, and then biofilm units were calculated. Results: Fourfold elevated glkA activity was observed upon the phosphorylation by BYK. Protein-protein docking analysis revealed that glkA structure docked close to the adenosine triphosphate-binding site of BYK structure corroborating the kinetic results. Further, S. aureus grown in the presence of elevated glucose concentration exhibited an increase in the rate of biofilm formation. Conclusion: The elevated function of glkA is an essential requirement for increased biofilm units in S. aureus, a key pathogenic factor that helps its survival and spread the infection. |
| نویسندگان مقاله |
dudipeta vasu | dudipeta vasu
department of biotechnology, sri venkateswara institute of medical sciences, tirupati 517 507, andhra pradesh, india
pasupuleti santhosh کومار | pasupuleti santhosh kumar
department of biotechnology, sri venkateswara institute of medical sciences, tirupati 517 507, andhra pradesh, india
uppu venkateswara prasad | uppu venkateswara prasad
department of biotechnology, sri venkateswara institute of medical sciences, tirupati 517 507, andhra pradesh, india
vimjam swarupa | vimjam swarupa
department of biotechnology, sri venkateswara institute of medical sciences, tirupati 517 507, andhra pradesh, india
sthanikam yeswanth | sthanikam yeswanth
department of biotechnology, sri venkateswara institute of medical sciences, tirupati 517 507, andhra pradesh, india
lokanathan srikanth | lokanathan srikanth
department of biotechnology, sri venkateswara institute of medical sciences, tirupati 517 507, andhra pradesh, india
manne mudhu sunitha | manne mudhu sunitha
department of biotechnology, sri venkateswara institute of medical sciences, tirupati 517 507, andhra pradesh, india
abhijith choudhary | abhijith choudhary
department of microbiology, sri venkateswara institute of medical sciences, tirupati 517 507, andhra pradesh, india
potukuchi venkata gurunadha krishna sarma | potukuchi venkata gurunadha krishna sarma
department of biotechnology, sri venkateswara institute of medical sciences, tirupati 517 507, andhra pradesh, india
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