Molecular modeling of peptide antibiotics hPAB-?and its artificial mutants construction
- VernacularTitle:人肽抗生素hPAB-?分子的同源模建及其突变体设计
- Author:
Xiancai RAO
;
Xiaolin JIN
;
Shu LI
;
Jinchuan HU
;
Xiaomei HU
;
Zhijin CHEN
;
Fuquan HU
;
- Publication Type:Journal Article
- Keywords:
Peptide antibiotics;
Homology modeling;
Mutants
- From:
Journal of Medical Postgraduates
2003;0(09):-
- CountryChina
- Language:Chinese
-
Abstract:
Objectives: To design the mutants of peptide antibiotics hPAB ? based on its molecular structure. Methods: The three dimension structure of hPAB ? was constructed by protein homology modeling method. The mutant molecules were designed and generated by PCR and inserted into pQE CP4 expression plasmid. The recombinant plasmids were identified by PCR and DNA sequencing and then transformed into Escherichia coli JM109 to express target fusion proteins. Results:Peptide hPAB ? shows one ? helical and three ? sheet in its structure. Its ? helical regions seem play a key role in the formation of active oligomer. Aside from positioning Thr 7 and Lys 10 into contact positions, the orientation of the ? helix is conserved about the oligome core, forming a ridge around it. Additionally, the dipoles of the helices would overlap to create a positively charged region near the core. These dipoles may be offset, however, by the presence of Asp 4 at the base of the helix. Two mutant molecules, hPAB ? 38 and hPAB ? 34, were designed by deleting N or/and C terminal 2～5 amino acid residues based on hPAB ? structure. The recombinant plasmids containing the mutants gene can express interest fusion proteins in E. coli JM109 successfully. Conclusions: Design, cloning and expression of the mutants of peptide antibiotics hPAB ? lay down the foundation for screening of the mutant of shorter peptide chain and having high or same antimicrobial activity.
