Abstract
We demonstrated lanthionine introduction into hexa-histidine-tagged (His-tagged) nukacin ISK-1 prepeptide NukA by modification enzyme NukM in Escherichia coli. Co-expression of nukA and nukM, purification of the resulting His-tagged prepeptide by affinity chromatography, and subsequent mass spectrometry analysis showed that the prepeptide was converted into a postulated peptide with decrease in mass of 72 Da which resulted from dehydration of four amino acids. Characterization of the resultant prepeptide indicated the presence of unusual amino acids, such as dehydrated amino acid, lanthionine or 3-methyllanthionine, in its C-terminal propeptide moiety. The modified prepeptide encompassing the leader peptide attached to the post-translationally modified propeptide moiety was readily obtained by one-step purification. Our findings will thus be a powerful tool for introducing unusual amino acids aimed at peptide engineering and also helpful to provide new insight for further understanding of lanthionine-forming enzymes for lantibiotics.
| Original language | English |
|---|---|
| Pages (from-to) | 507-513 |
| Number of pages | 7 |
| Journal | Biochemical and Biophysical Research Communications |
| Volume | 336 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - Oct 21 2005 |
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All Science Journal Classification (ASJC) codes
- Biochemistry
- Biophysics
- Molecular Biology
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Lanthionine introduction into nukacin ISK-1 prepeptide by co-expression with modification enzyme NukM in Escherichia coli. / Nagao, Jun Ichi; Harada, Yoshitaka; Shioya, Kouki; Aso, Yuji; Zendo, Takeshi; Nakayama, Jiro; Sonomoto, Kenji.
In: Biochemical and Biophysical Research Communications, Vol. 336, No. 2, 21.10.2005, p. 507-513.Research output: Contribution to journal › Article
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TY - JOUR
T1 - Lanthionine introduction into nukacin ISK-1 prepeptide by co-expression with modification enzyme NukM in Escherichia coli
AU - Nagao, Jun Ichi
AU - Harada, Yoshitaka
AU - Shioya, Kouki
AU - Aso, Yuji
AU - Zendo, Takeshi
AU - Nakayama, Jiro
AU - Sonomoto, Kenji
PY - 2005/10/21
Y1 - 2005/10/21
N2 - We demonstrated lanthionine introduction into hexa-histidine-tagged (His-tagged) nukacin ISK-1 prepeptide NukA by modification enzyme NukM in Escherichia coli. Co-expression of nukA and nukM, purification of the resulting His-tagged prepeptide by affinity chromatography, and subsequent mass spectrometry analysis showed that the prepeptide was converted into a postulated peptide with decrease in mass of 72 Da which resulted from dehydration of four amino acids. Characterization of the resultant prepeptide indicated the presence of unusual amino acids, such as dehydrated amino acid, lanthionine or 3-methyllanthionine, in its C-terminal propeptide moiety. The modified prepeptide encompassing the leader peptide attached to the post-translationally modified propeptide moiety was readily obtained by one-step purification. Our findings will thus be a powerful tool for introducing unusual amino acids aimed at peptide engineering and also helpful to provide new insight for further understanding of lanthionine-forming enzymes for lantibiotics.
AB - We demonstrated lanthionine introduction into hexa-histidine-tagged (His-tagged) nukacin ISK-1 prepeptide NukA by modification enzyme NukM in Escherichia coli. Co-expression of nukA and nukM, purification of the resulting His-tagged prepeptide by affinity chromatography, and subsequent mass spectrometry analysis showed that the prepeptide was converted into a postulated peptide with decrease in mass of 72 Da which resulted from dehydration of four amino acids. Characterization of the resultant prepeptide indicated the presence of unusual amino acids, such as dehydrated amino acid, lanthionine or 3-methyllanthionine, in its C-terminal propeptide moiety. The modified prepeptide encompassing the leader peptide attached to the post-translationally modified propeptide moiety was readily obtained by one-step purification. Our findings will thus be a powerful tool for introducing unusual amino acids aimed at peptide engineering and also helpful to provide new insight for further understanding of lanthionine-forming enzymes for lantibiotics.
UR - http://www.scopus.com/inward/record.url?scp=24644468692&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=24644468692&partnerID=8YFLogxK
U2 - 10.1016/j.bbrc.2005.08.125
DO - 10.1016/j.bbrc.2005.08.125
M3 - Article
C2 - 16143300
AN - SCOPUS:24644468692
VL - 336
SP - 507
EP - 513
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 2
ER -