Gamma-phage lysin PlyG sequence-based synthetic peptides coupled with Qdot-nanocrystals are useful for developing detection methods for Bacillus anthracis by using its surrogates, B. anthracis-Sterne and B. cereus-4342

Previous reports of site-directed deletion analysis on gamma (gamma)-phage lysin protein (PlyG) have demonstrated that removal of a short amino acid sequence in the C-terminal region encompassing a 10-amino acid motif (190LKMTADFILQ199) abrogates its binding activity specific to the cell wall of Bac...

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Published in:	BMC biotechnology Vol. 9; no. 1; p. 67
Main Authors:	Sainathrao, Shilpakala, Mohan, Ketha V Krishna, Atreya, Chintamani
Format:	Journal Article
Language:	English
Published:	England BioMed Central Ltd 22-07-2009 BioMed Central BMC
Subjects:	Amino Acid Sequence Anthrax Anthrax - diagnosis Anthrax - microbiology Bacillus anthracis Bacillus anthracis - isolation & purification Bacillus cereus Bacillus Phages Bacterial proteins Bacterial vaccines BASIC BIOLOGICAL SCIENCES Binding Sites Biotechnology & applied microbiology Cell Wall - microbiology Diagnosis Enzyme-Linked Immunosorbent Assay high pressure liquid chromatography Molecular Sequence Data N-Acetylmuramoyl-L-alanine Amidase - metabolism Peptides - chemical synthesis Peptides - metabolism Physiological aspects Prevention Quantum Dots Sensitivity and Specificity spiked plasma synthetic peptide Viral Proteins - metabolism United States
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Summary:	Previous reports of site-directed deletion analysis on gamma (gamma)-phage lysin protein (PlyG) have demonstrated that removal of a short amino acid sequence in the C-terminal region encompassing a 10-amino acid motif (190LKMTADFILQ199) abrogates its binding activity specific to the cell wall of Bacillus anthracis. Whether short synthetic peptides representing the10-amino acid PlyG putative binding motif flanked by surrounding N- and C-terminal residues also selectively bind to the bacterial cell wall has not been evaluated. If such peptides do demonstrate selective binding to the cell wall, they could serve as bio-probes towards developing detection technologies for B. anthracis. Furthermore, by using B. anthracis (Sterne, 34F2), an animal vaccine and B. cereus-4342, a gamma-phage susceptible rare strain as surrogates of B. anthracis, development of proof-of-concepts for B. anthracis are feasible. Using four different methods, we evaluated six synthetic peptides representing the putative binding motif including flanking sequences (PlyG-P1 through P6) for the bacterial cell wall binding capacity. Our analysis identified PlyG-P1, PlyG-P3 and PlyG-P5 to have binding capability to both B. anthracis (Sterne, 34F2) and B. cereus-4342. The peptides however did not bind to B. cereus-11778, B. thuringiensis, and B. cereus-10876 suggesting their specificity for B. anthracis-Sterne and B. cereus-4342. PlyG-P3 in combination with fluorescent light microscopy detected even a single bacterium in plasma spiked with the bacteria. Overall, these studies illustrate that the short 10-amino acid sequence 'LKMTADFILQ' in fact is a stand-alone bacterial cell wall-binding motif of PlyG. In principle, synthetic peptides PlyG-P1, PlyG-P3 and PlyG-P5, especially PlyG-P3 coupled with Qdot-nanocrystals are useful as high-sensitivity bio-probes in developing detection technologies for B. anthracis.
Bibliography:	SC0014664 USDOE Office of Science (SC), Biological and Environmental Research (BER). Biological Systems Science Division
ISSN:	1472-6750 1472-6750
DOI:	10.1186/1472-6750-9-67