Reports

Disulfide Isomerization After Membrane Release of Its SAR Domain Activates P1 Lysozyme

Min Xu,^* Arockiasamy Arulandu,^* Douglas K. Struck, Stephanie Swanson, James C. Sacchettini, Ry Young

The P1 lysozyme Lyz is secreted to the periplasm of Escherichia coli and accumulates in an inactive membrane-tethered form. Genetic and biochemical experiments show that, when released from the bilayer, Lyz is activated by an intramolecular thiol-disulfide isomerization, which requires a cysteine in its N-terminal SAR (signal-arrest-release) domain. Crystal structures confirm the alternative disulfide linkages in the two forms of Lyz and reveal dramatic conformational differences in the catalytic domain. Thus, the exported P1 endolysin is kept inactive by three levels of control—topological, conformational, and covalent—until its release from the membrane is triggered by the P1 holin.

Department of Biochemistry and Biophysics, Texas A&M University, College Station, TX 77843–2128, USA.

^* These authors contributed equally to the results described in this work.

To whom correspondence should be addressed. E-mail: sacchett{at}tamu.edu (J.C.S.) and ryland{at}tamu.edu (R.Y.).

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Evolutionary dominance of holin lysis systems derives from superior genetic malleability.

Y. Zheng, D. K. Struck, C. A. Dankenbring, and R. Young (2008)
Microbiology 154, 1710-1718
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Molecular Control of Bacterial Death and Lysis.

K. C. Rice and K. W. Bayles (2008)
Microbiol. Mol. Biol. Rev. 72, 85-109
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The Pinholin of Lambdoid Phage 21: Control of Lysis by Membrane Depolarization.

T. Park, D. K. Struck, C. A. Dankenbring, and R. Young (2007)
J. Bacteriol. 189, 9135-9139
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The T4 RI Antiholin Has an N-Terminal Signal Anchor Release Domain That Targets It for Degradation by DegP.

T. A. T. Tran, D. K. Struck, and R. Young (2007)
J. Bacteriol. 189, 7618-7625
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Topological dynamics of holins in programmed bacterial lysis.

T. Park, D. K. Struck, J. F. Deaton, and R. Young (2006)
PNAS 103, 19713-19718
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Genomic Analysis of Pseudomonas aeruginosa Phages LKD16 and LKA1: Establishment of the {phi}KMV Subgroup within the T7 Supergroup..

P.-J. Ceyssens, R. Lavigne, W. Mattheus, A. Chibeu, K. Hertveldt, J. Mast, J. Robben, and G. Volckaert (2006)
J. Bacteriol. 188, 6924-6931
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Bacteriophage endolysins as a novel class of antibacterial agents..

J. Borysowski, B. Weber-Dabrowska, and A. Gorski (2006)
Experimental Biology and Medicine 231, 366-377
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Divergence and Mosaicism among Virulent Soil Phages of the Burkholderia cepacia Complex.

E. J. Summer, C. F. Gonzalez, M. Bomer, T. Carlile, A. Embry, A. M. Kucherka, J. Lee, L. Mebane, W. C. Morrison, L. Mark, et al. (2006)
J. Bacteriol. 188, 255-268
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Lysis Timing and Bacteriophage Fitness.

I.-N. Wang (2006)
Genetics 172, 17-26
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COG3926 and COG5526: A tale of two new lysozyme-like protein families.

J. Pei and N. V. Grishin (2005)
Protein Sci. 14, 2574-2581
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