DOI: 10.1126/science.1092600. Lacticin 481: In Vitro Reconstitution of Lantibiotic Synthetase Activity

Lacticin 481: In Vitro Reconstitution of Lantibiotic Synthetase Activity

10.1126/science.1092600

Crossref journal-article

American Association for the Advancement of Science (AAAS)

Science (221)

Abstract

The lantibiotic lacticin 481 is synthesized on ribosomes as a prepeptide (LctA) and posttranslationally modified to its mature form. These modifications include dehydration of serines and threonines, followed by intramolecular addition of cysteines to the unsaturated amino acids, which generates cyclic thioethers. This process breaks eight chemical bonds and forms six newbonds and is catalyzed by one enzyme, LctM. We have characterized the in vitro activity of LctM, which completely processed a series of LctA mutants, displaying a permissive substrate specificity that holds promise for antibiotic engineering.

Bibliography

Xie, L., Miller, L. M., Chatterjee, C., Averin, O., Kelleher, N. L., & van der Donk, W. A. (2004). Lacticin 481: In Vitro Reconstitution of Lantibiotic Synthetase Activity. Science, 303(5658), 679â681.

Authors 6

Lili Xie (first)
Leah M. Miller (additional)
Champak Chatterjee (additional)
Olga Averin (additional)
Neil L. Kelleher (additional)
Wilfred A. van der Donk (additional)

References 29 Referenced 211

10.1126/science.286.5448.2361
T. R. Klaenhammer, FEMS Microbiol. Rev.12, 39 (1993). (10.1016/0168-6445(93)90057-G) / FEMS Microbiol. Rev. (1993)
H. G. Sahl, G. Bierbaum, Annu. Rev. Microbiol.52, 41 (1998). (10.1146/annurev.micro.52.1.41) / Annu. Rev. Microbiol. (1998)
H. W. van den Hoovenet al., FEBS Lett.391, 317 (1996). (10.1016/0014-5793(96)00771-5) / FEBS Lett. (1996)
10.1007/BF00399424
W. Liu, J. N. Hansen, J. Biol. Chem.267, 25078 (1992). (10.1016/S0021-9258(19)74008-3) / J. Biol. Chem. (1992)
O. P. Kuiperset al., Antonie Van Leeuwenhoek69, 161 (1996). (10.1007/BF00399421) / Antonie Van Leeuwenhoek (1996)
P. Chenet al., Appl. Environ. Microbiol.64, 2335 (1998). (10.1128/AEM.64.7.2335-2340.1998) / Appl. Environ. Microbiol. (1998)
C. Szekat, R. W. Jack, D. Skutlarek, H. Farber, G. Bierbaum, Appl. Environ. Microbiol.69, 3777 (2003). (10.1128/AEM.69.7.3777-3783.2003) / Appl. Environ. Microbiol. (2003)
Lack of lantibiotic production may be due to disruption of the posttranslational modification process degradation of the non-native peptide product or intermediates breakdown of self-immunity resulting in shutdown of production or perturbation of signaling pathways in cases in which the bacteriocin acts as a quorum sensor controlling its own expression. See references ( 25 â 28 ).
N. Schnellet al., Nature333, 276 (1988). (10.1038/333276a0) / Nature (1988)
A. RinceÌ, A. Dufour, P. Uguen, J. P. Le Pennec, D. Haras, Appl. Environ. Microbiol.63, 4252 (1997). (10.1128/aem.63.11.4252-4260.1997) / Appl. Environ. Microbiol. (1997)
J. C. Piard, O. P. Kuipers, H. S. Rollema, M. J. Desmazeaud, W. M. de Vos, J. Biol. Chem.268, 16361 (1993). (10.1016/S0021-9258(19)85429-7) / J. Biol. Chem. (1993)
O. Koponenet al., Microbiol.148, 3561 (2002). (10.1099/00221287-148-11-3561) / Microbiol. (2002)
10.1007/BF00399422
Materials and methods are available as supporting material on Science Online.
The sequence of the His 6 -tag linker is GSSHHHHHHSSGLVPRGSH. The DNA sequences of all constructs were verified and the masses of the expressed gene products were determined by mass spectrometry.
10.1021/bi0354942
10.1126/science.284.5418.1289
P. Roepstorff, J. Fohlman, Biomed. Mass Spectrom.11, 601 (1984). (10.1002/bms.1200111109) / Biomed. Mass Spectrom. (1984)
A. J. Kleinnijenhuis, M. C. Duursma, E. Breuking, R. M. A. Heeren, A. J. R. Heck, Anal. Chem.75, 3219 (2003). (10.1021/ac0263770) / Anal. Chem. (2003)
10.1126/science.274.5290.1188
T. W. Muir, D. Sondhi, P. A. Cole, Proc. Natl. Acad. Sci. U.S.A.95, 6705 (1998). (10.1073/pnas.95.12.6705) / Proc. Natl. Acad. Sci. U.S.A. (1998)
L. Xie thesis University of Illinois Urbana-Champaign (2003).
J. R. van der Meeret al., J. Biol. Chem.269, 3555 (1994). (10.1016/S0021-9258(17)41899-0) / J. Biol. Chem. (1994)
O. P. Kuipers, M. M. Beerthuyzen, P. G. de Ruyter, E. J. Luesink, W. M. de Vos, J. Biol. Chem.270, 27299 (1995). (10.1074/jbc.270.45.27299) / J. Biol. Chem. (1995)
M. Reis, M. Eschbach-Bludau, M. I. Iglesias-Wind, T. Kupke, H. G. Sahl, Appl. Environ. Microbiol.60, 2876 (1994). (10.1128/aem.60.8.2876-2883.1994) / Appl. Environ. Microbiol. (1994)
G. Bierbaum, M. Reis, C. Szekat, H. G. Sahl, Appl. Environ. Microbiol.60, 4332 (1994). (10.1128/aem.60.12.4332-4338.1994) / Appl. Environ. Microbiol. (1994)
This work was supported by grants from the National Institutes of Health to W.A.V. (GM58822) and N.L.K. (GM 067725) the Beckman Foundation and the Burroughs Wellcome Fund (APP 1920 to W.A.V.).

Dates

Type	When
Created	21 years, 7 months ago (Jan. 29, 2004, 5:58 p.m.)
Deposited	1 year, 7 months ago (Jan. 9, 2024, 10:51 p.m.)
Indexed	1 month, 1 week ago (July 24, 2025, 8:03 a.m.)
Issued	21 years, 7 months ago (Jan. 30, 2004)
Published	21 years, 7 months ago (Jan. 30, 2004)
Published Print	21 years, 7 months ago (Jan. 30, 2004)

Funders 0

None

BibTeX

@article{Xie_2004, title={Lacticin 481: In Vitro Reconstitution of Lantibiotic Synthetase Activity}, volume={303}, ISSN={1095-9203}, url={http://dx.doi.org/10.1126/science.1092600}, DOI={10.1126/science.1092600}, number={5658}, journal={Science}, publisher={American Association for the Advancement of Science (AAAS)}, author={Xie, Lili and Miller, Leah M. and Chatterjee, Champak and Averin, Olga and Kelleher, Neil L. and van der Donk, Wilfred A.}, year={2004}, month=jan, pages={679–681} }

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