Expression, Purification, and Functional Characterization of Atypical Xenocin, Its Immunity Protein, and Their Domains from Xenorhabdus nematophila
Author
Source
International Journal of Bacteriology
Issue
Vol. 2013, Issue 2013 (31 Dec. 2013), pp.1-9, 9 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2013-12-18
Country of Publication
Egypt
No. of Pages
9
Main Subjects
Abstract EN
Xenorhabdus nematophila, a gram-negative bacterium belonging to the family Enterobacteriaceae is a natural symbiont of a soil nematode from the family Steinernematidae.
In this study cloning, expression, and purification of broad range iron regulated multidomain bacteriocin called xenocin from X.
nematophila (66 kDa, encoded by xcinA gene) and its multidomain immunity protein (42 kDa, encoded by ximB gene) have been done.
xcinA-ximB (N′ terminal 270 bp), translocation, and translocation-receptor domain of xcinA, ximB, and its hemolysin domain were cloned, expressed, and purified by single step Ni-NTA chromatography under native conditions.
In the functional characterization, neutralization of xcinA toxicity by immunity domain of ximB gene was determined by endogenous assay.
Exogenous toxic assays results showed that only the purified recombinant xenocin-immunity domain (10 kDa) protein complex had toxic activity.
Atypical cognate immunity protein (42 kDa) of xenocin was fusion of immunity domain (10 kDa) and hemolysin domain (32 kDa).
In silico analysis of immunity protein revealed its similarity with hemolysin and purine NTPase like proteins.
Hemolytic activity was not observed in immunity protein or in its various domains; however, full-length immunity protein lacking Walker motif showed ATPase activity.
Finally, using circular dichroism performed secondary structural analyses of all the recombinant proteins/protein complexes.
American Psychological Association (APA)
Rathore, Jitendra Singh. 2013. Expression, Purification, and Functional Characterization of Atypical Xenocin, Its Immunity Protein, and Their Domains from Xenorhabdus nematophila. International Journal of Bacteriology،Vol. 2013, no. 2013, pp.1-9.
https://search.emarefa.net/detail/BIM-495452
Modern Language Association (MLA)
Rathore, Jitendra Singh. Expression, Purification, and Functional Characterization of Atypical Xenocin, Its Immunity Protein, and Their Domains from Xenorhabdus nematophila. International Journal of Bacteriology No. 2013 (2013), pp.1-9.
https://search.emarefa.net/detail/BIM-495452
American Medical Association (AMA)
Rathore, Jitendra Singh. Expression, Purification, and Functional Characterization of Atypical Xenocin, Its Immunity Protein, and Their Domains from Xenorhabdus nematophila. International Journal of Bacteriology. 2013. Vol. 2013, no. 2013, pp.1-9.
https://search.emarefa.net/detail/BIM-495452
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-495452