Please use this identifier to cite or link to this item:
|Title:||Identification of the essential histidine residue for high-affinity binding of AlbA protein to albicidin antibiotics||Authors:||Weng, L.-X.
|Issue Date:||1-Feb-2003||Citation:||Weng, L.-X., Xu, J.-L., Li, Q., Birch, R.G., Zhang, L.-H. (2003-02-01). Identification of the essential histidine residue for high-affinity binding of AlbA protein to albicidin antibiotics. Microbiology 149 (2) : 451-457. ScholarBank@NUS Repository. https://doi.org/10.1099/mic.0.25942-0||Abstract:||The albA gene from Klebsiella oxytoca encodes a protein that binds albicidin phytotoxins and antibiotics with high affinity. Previously, it has been shown that shifting pH from 6 to 4 reduces binding activity of AlbA by about 30 %, indicating that histidine residues might be involved in substrate binding. In this study, molecular analysis of the albA coding region revealed sequence discrepancies with the albA sequence reported previously, which were probably due to sequencing errors. The albA gene was subsequently cloned from K. oxytoca ATCC 13182T to establish the revised sequence. Biochemical and molecular approaches were used to determine the functional role of four histidine residues (His78, His125, His141 and His189) in the corrected sequence for AlbA. Treatment of AlbA with diethyl pyrocarbonate (DEPC), a histidine-specific alkylating reagent, reduced binding activity by about 95 %. DEPC treatment increased absorbance at 240-244 nm by an amount indicating conversion to N-carbethoxyhistidine of a single histidine residue per AlbA molecule. Pretreatment with albicidin protected AlbA against modification by DEPC, with a 1:1 molar ratio of albicidin to the protected histidine residues. Based on protein secondary structure and amino acid surface probability indices, it is predicted that His125 might be the residue required for albicidin binding. Mutation of His125 to either alanine or leucine resulted in about 32 % loss of binding activity, and deletion of His125 totally abolished binding activity. Mutation of His125 to arginine and tyrosine had no effect. These results indicate that His125 plays a key role either in an electrostatic interaction between AlbA and albicidin or in the conformational dynamics of the albicidin-binding site.||Source Title:||Microbiology||URI:||http://scholarbank.nus.edu.sg/handle/10635/102467||ISSN:||13500872||DOI:||10.1099/mic.0.25942-0|
|Appears in Collections:||Staff Publications|
Show full item record
Files in This Item:
There are no files associated with this item.
checked on Feb 20, 2020
WEB OF SCIENCETM
checked on Feb 20, 2020
checked on Feb 16, 2020
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.