Biochemical and physical properties for a recombinant IL6 Pseudomonas exotoxin fusion protein IL6D24-PE40KDEL.
- Author:
Jian-Wu CUI
1
;
Si-Qi GUO
;
Yu-Ying SUN
;
Nan LIU
;
Fei LIANG
;
Yong-Zhi XI
Author Information
1. Department of Immunology, Affiliated Hospital of Academy of Military Medical Sciences, Beijing, China.
- Publication Type:Journal Article
- MeSH:
ADP Ribose Transferases;
chemistry;
metabolism;
pharmacology;
Amino Acid Sequence;
Blotting, Western;
Cell Line, Tumor;
Cell Survival;
drug effects;
Dose-Response Relationship, Drug;
Exotoxins;
chemistry;
metabolism;
pharmacology;
Humans;
Interleukin-6;
chemistry;
metabolism;
pharmacology;
Molecular Sequence Data;
Pseudomonas aeruginosa;
genetics;
metabolism;
Recombinant Fusion Proteins;
chemistry;
metabolism;
pharmacology;
Sequence Analysis, Protein;
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
- From:
Journal of Experimental Hematology
2004;12(6):825-828
- CountryChina
- Language:Chinese
-
Abstract:
The objective was to identify some biochemical and physical properties for fusion protein IL6D24-PE40KDEL. Edman degradation, SDS-PAGE, peptide mass fingerprinting, Western blot and MTT were used for identification of the protein. The results showed that the sequence of N-terminus is Met-Ile-Asp-Lys-Gln-Ile, Met was added because of prokaryotic expression system; Western blot revealed that the purified protein could react with IL6 and PEA antibody. The purified protein IL6D24-PE40KDEL could kill the multiple myeloma cell lines U266 expressing high affinity IL6R, but it could not kill the cell lines CEM which not expressed IL6R; The molecular weight was 58.7 kD measuring by SDS-PAGE; peptide mass fingerprinting (PMF) confirmed that the construction of IL6D24-PE40KDEL was correct. A novel protein by Peptident database in EXPASY web site was identified. In conclusion, IL6D24-PE40KDEL is a new targeting protein with bioactivity of specific killing effect.
