Mass spectrometric sequencing of endotoxin proteins of Bacillus thuringiensis ssp. konkukian extracted from polyacrylamide gels

The amino acid sequences of the crystal proteins of Bacillus thuringiensis ssp. konkukian strain HL‐47 are unknown. We used 1‐D denaturing polyacrylamide electrophoresis, nano‐ESI‐Q‐TOF‐MS, and protein database searching to analyze these proteins. On SDS‐PAGE gels, a preparation of purified crystal...

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Bibliographic Details
Published in:Proteomics (Weinheim) 2006-03, Vol.6 (5), p.1512-1517
Main Authors: Lee, Kwang Yong, Kang, Eun Young, Park, Sooil, Ahn, Seong Kyu, Yoo, Kwan Hee, Kim, Jin Young, Lee, Hyung Hoan
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Analytical, structural and metabolic biochemistry
Bacillus thuringiensis
Bacterial Proteins - chemistry
Bacterial Proteins - genetics
Bacterial Toxins - chemistry
Bacterial Toxins - genetics
Biological and medical sciences
Electrophoresis, Polyacrylamide Gel
Endotoxin crystal
Endotoxins - chemistry
Endotoxins - genetics
ESI-Q-TOF mass spectrometry
Fundamental and applied biological sciences. Psychology
Hemolysin Proteins
Insecticides - chemistry
Mass Spectrometry
Miscellaneous
Molecular Sequence Data
Peptides - chemistry
Peptides - genetics
Proteins
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Summary:The amino acid sequences of the crystal proteins of Bacillus thuringiensis ssp. konkukian strain HL‐47 are unknown. We used 1‐D denaturing polyacrylamide electrophoresis, nano‐ESI‐Q‐TOF‐MS, and protein database searching to analyze these proteins. On SDS‐PAGE gels, a preparation of purified crystal proteins exhibited 110, 102, 76, 55, 37, and 30 kDa protein bands. Immunoblotting of the gel with antiserum raised to this preparation revealed that four crystal proteins, of 110, 102, 55, and 37 kDa, reacted with the specific antiserum. The 102‐kDa major protein reacted strongly. The other crystal proteins showed weak immunoreactivity. The 102 and 55 kDa proteins were analyzed by ESI‐MS. The internal amino acid sequence of the 102‐kDa major protein has similarity to the sequences of the surface layer protein of B. thuringiensis ssp. finitimus and B. anthracis. However, the internal amino acid sequences of the 55 kDa protein did not show any homology to proteins in the databases. Proteomic analysis of these proteins leads to the conclusion that the sequence data provided the protein databases of the crystal proteins of the konkukian ssp.
ISSN:1615-9853
1615-9861
DOI:10.1002/pmic.200500298