TAILIEUCHUNG - Investigation of the in vivo interaction between β-lactamase and its inhibitor protein
The affinity of β-lactamase inhibitory protein (BLIP) for TEM-1 β-lactamase has raised hopes in the challenge of proteinbased inhibitor discovery for β-lactamase-mediated antibiotic resistance. Currently, the effect of the formation of the β-lactamase:BLIP complex in vivo in β-lactam resistant bacteria is an open question. | Turkish Journal of Biology Turk J Biol (2015) 39: 485-492 © TÜBİTAK doi: Research Article Investigation of the in vivo interaction between β-lactamase and its inhibitor protein 1 2 1 1 Nilay BÜDEYRİ GÖKGÖZ , Simay YALAZ , Naze Gül AVCI , Gizem BULDUM , 2 1, Elif ÖZKIRIMLI ÖLMEZ , Berna SARIYAR AKBULUT * 1 Bioengineering Department, Marmara University, Göztepe Campus, Kadıköy, İstanbul, Turkey 2 Chemical Engineering Department, Boğaziçi University, Bebek, İstanbul, Turkey Received: Accepted/Published Online: Printed: Abstract: The affinity of β-lactamase inhibitory protein (BLIP) for TEM-1 β-lactamase has raised hopes in the challenge of proteinbased inhibitor discovery for β-lactamase-mediated antibiotic resistance. Currently, the effect of the formation of the β-lactamase:BLIP complex in vivo in β-lactam resistant bacteria is an open question. The scarcity of information to the extent to which BLIP can impair β-lactamase activity inside cells has urged us to assess the in vivo efficacy of BLIP as a potent β-lactamase inhibitor. To this end, β-lactamase and BLIP were coexpressed in Escherichia coli. Simultaneous expression of β-lactamase and BLIP and the formation of the TEM-1 β-lactamase:BLIP complex in the periplasmic space of E. coli were verified by electrophoretic and Western blot techniques. Growth profiles of the cells expressing both β-lactamase and its protein inhibitor, complemented with β-lactamase activity measurements, suggested that BLIP synthesis retarded cell growth and reduced β-lactamase activity. Although co-expression of β-lactamase and its protein inhibitor did not completely impair cell growth, the specificity of BLIP enabled it to bind β-lactamase in the bacterial periplasm, regardless of the crowding components. Key words: β-lactamase, antibiotic resistance, BLIP 1. Introduction β-lactam antibiotics hold great importance in healing diverse
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