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Mass spectrometric sequencing of endotoxin proteins of Bacillus thuringiensis ssp. konkukian extracted from polyacrylamide gels
Author(s) -
Lee Kwang Yong,
Kang Eun Young,
Park Sooil,
Ahn Seong Kyu,
Yoo Kwan Hee,
Kim Jin Young,
Lee Hyung Hoan
Publication year - 2006
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200500298
Subject(s) - bacillus thuringiensis , amino acid , gel electrophoresis , homology (biology) , peptide sequence , biochemistry , polyacrylamide gel electrophoresis , biology , molecular mass , antiserum , microbiology and biotechnology , protein sequencing , chemistry , bacteria , gene , antibody , enzyme , genetics
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.