SavedAdd your journal

Journals / Turkish Journal of Biology / 2015 / Cilt: 39 - Sayı: 3

Investigation of the in vivo interaction between ß-lactamase and its inhibitor protein

Nilay Büdeyri GÖKGÖZ
Simay YALAZ
Naze Gül AVCI
Gizem BULDUM
Elif Özkirimli ÖLMEZ
Berna Sariyar AKBULUT
Pages
485–492
Publication Date
2015-08-13
DOI
—
🏛️Bibliographic metadata and full text access for this article are provided via DergiPark.
DergiPark Record ↗
✨

AI Summary & Key Takeaways

AI Insight
⚡ TL;DR(Too Long; Didn't Read)

Although co-expression of ß-lactamase and its protein inhibitor did not completely impair cell growth, the specificity of BLIP enabled it to bind ß-lactamase in the bacterial periplasm, regardless of the crowding components.

📌 3 Key Takeaways

🎯
Objective & Hypothesis:The affinity of ß-lactamase inhibitory protein (BLIP) for TEM-1 ß-lactamase has raised hopes in the challenge of protein-based inhibitor discovery for ß-lactamase-mediated antibiotic resistance.
📊
Key Findings & Data:Simultaneous expression of ß-lactamase and BLIP and the formation of the TEM-1 ß-lactamase:BLIP complex in the periplasmic space of E. coli were verified by electrophoretic and Western blot techniques.
💡
Conclusion & Impact:Although co-expression of ß-lactamase and its protein inhibitor did not completely impair cell growth, the specificity of BLIP enabled it to bind ß-lactamase in the bacterial periplasm, regardless of the crowding components.
🔬Design:Research Article
📍Scope:Comprehensive Sample
⚙️Variables:ß-lactamase · antibiotic resistance · BLIP

Abstract

The affinity of ß-lactamase inhibitory protein (BLIP) for TEM-1 ß-lactamase has raised hopes in the challenge of protein-based inhibitor discovery for ß-lactamase-mediated antibiotic resistance. Currently, the effect of the formation of the ß-lactamase:BLIP complex in vivo in ß-lactam resistant bacteria is an open question. The scarcity of information to the extent to which BLIP can impair ß-lactamase activity inside cells has urged us to assess the in vivo efficacy of BLIP as a potent ß-lactamase inhibitor. To this end, ß-lactamase and BLIP were coexpressed in Escherichia coli. Simultaneous expression of ß-lactamase and BLIP and the formation of the TEM-1 ß-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 ß-lactamase and its protein inhibitor, complemented with ß-lactamase activity measurements, suggested that BLIP synthesis retarded cell growth and reduced ß-lactamase activity. Although co-expression of ß-lactamase and its protein inhibitor did not completely impair cell growth, the specificity of BLIP enabled it to bind ß-lactamase in the bacterial periplasm, regardless of the crowding components.

Özet

The affinity of ß-lactamase inhibitory protein (BLIP) for TEM-1 ß-lactamase has raised hopes in the challenge of protein-based inhibitor discovery for ß-lactamase-mediated antibiotic resistance. Currently, the effect of the formation of the ß-lactamase:BLIP complex in vivo in ß-lactam resistant bacteria is an open question. The scarcity of information to the extent to which BLIP can impair ß-lactamase activity inside cells has urged us to assess the in vivo efficacy of BLIP as a potent ß-lactamase inhibitor. To this end, ß-lactamase and BLIP were coexpressed in Escherichia coli. Simultaneous expression of ß-lactamase and BLIP and the formation of the TEM-1 ß-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 ß-lactamase and its protein inhibitor, complemented with ß-lactamase activity measurements, suggested that BLIP synthesis retarded cell growth and reduced ß-lactamase activity. Although co-expression of ß-lactamase and its protein inhibitor did not completely impair cell growth, the specificity of BLIP enabled it to bind ß-lactamase in the bacterial periplasm, regardless of the crowding components.

🔖

Cite This Article

GÖKGÖZ, N. B., YALAZ, S., AVCI, N. G., BULDUM, G., ÖLMEZ, E. Ö., AKBULUT, B. S. (2015). Investigation of the in vivo interaction between ß-lactamase and its inhibitor protein. Turkish Journal of Biology, Cilt: 39 - Sayı: 3, 485–492. https://ajindex.com/articles/181759

📄 Full Text PDF Viewer

📄

PDF görüntüleyici hazırlanıyor...

References (12)

  1. Adelowo OO, Fagade OE (2012). Phylogenetic characterization, antimicrobial susceptibilities, and mechanisms of resistance in bacteria isolates from a poultry waste-polluted river, southwestern Nigeria. Turk J Biol 36: 37–45.
  2. Alaybeyoglu B, Sariyar Akbulut B, Ozkirimli E (2015). A novel chimeric peptide with antimicrobial activity. J Pept Sci, accepted for publication. Bebrone C, Moali C, Mahy F, Rival S, Docquier JD, Rossolini GM, Fastrez J, Pratt RF, Frere JM, Galleni M (2001). CENTA as a chromogenic substrate for studying beta-lactamases. Antimicrob Agents Ch 45: 1868–1871.
  3. Blakesley RW, Boezi JA (1977). A new staining technique for proteins in polyacrylamide gels using Coomassie Brilliant Blue G250. Anal Biochem 82: 580–582.
  4. Blazquez J, Baquero MR, Canton R, Alos I, Baquero F (1993). Characterization of a new TEM-type β-lactamase resistant to clavulanate, sulbactam, and tazobactam in a clinical isolate of Escherichia coli. Antimicrob Agents Ch 37: 2059–2063.
  5. Bradford MM (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254.
  6. Chanda S, Rakholiya K, Dholakia K, Baravalia Y (2013). Antimicrobial, antioxidant, and synergistic properties of two nutraceutical plants: Terminalia catappa L. and Colocasia esculenta L. Turk J Biol 37: 81–91.
  7. Doran JL, Leskiw BK, Aippersbach S, Jensen SE (1990). Isolation and characterization of a beta-lactamase-inhibitory protein from Streptomyces clavuligerus and clonning and analysis of the corresponding gene. J Bacteriol 172: 4909–4918.
  8. Essack SY (2001). The development of beta-lactam antibiotics in response to the evolution of beta-lactamases. Pharm Res 18: 1391–1399. Frère JM (1995). Beta-lactamases and bacterial resistance to antibiotics. Mol Microbiol 16: 385–395.
  9. Fryszczyn BG, Brown NG, Huang W, Balderas MA, Palzkill T (2011). Use of periplasmic target protein capture for phage display engineering of tight-binding protein-protein interactions. Protein Eng Des Sel 24: 819–828.
  10. Hanes MS, Ratcliff K, Marqusee S, Handel TM (2010). Protein- protein binding affinities by pulse proteolysis: application to TEM-1/BLIP protein complexes. Protein Sci 19: 1996–2000.
  11. Huang W, Petrosino J, Hirsch M, Shenkin PS, Palzkill T (1996). Amino acid sequence determinants of β-lactamase structure and activity. J Mol Biol 258: 688–703.
  12. Huang W, Beharry Z, Zhang Z, Palzkill T (2003). A broad-spectrum peptide inhibitor of beta-lactamase id