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Journal of Bacteriology, October 1999, p. 6306-6311, Vol. 181, No. 20
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
In Vitro Characterization of
Peptidoglycan-Associated Lipoprotein (PAL)-Peptidoglycan and
PAL-TolB Interactions
Emmanuelle
Bouveret,*
Hélène
Bénédetti,
Alain
Rigal,
Erwann
Loret, and
Claude
Lazdunski
Laboratoire d'Ingéniérie des
Systèmes Macromoléculaires, Institut de Biologie
Structurale et Microbiologie, Centre National de la Recherche
Scientifique (CNRS), 13402 Marseille Cedex 20, France
Received 6 May 1999/Accepted 26 July 1999
The Tol-peptidoglycan-associated lipoprotein (PAL) system of
Escherichia coli is a multiprotein complex of the envelope
involved in maintaining outer membrane integrity. PAL and the
periplasmic protein TolB, two components of this complex, are
interacting with each other, and they have also been reported to
interact with OmpA and the major lipoprotein, two proteins interacting with the peptidoglycan. All these interactions suggest a role of the
Tol-PAL system in anchoring the outer membrane to the peptidoglycan. Therefore, we were interested in better understanding the interaction between PAL and the peptidoglycan. We designed an in vitro interaction assay based on the property of purified peptidoglycan to be pelleted by
ultracentrifugation. Using this assay, we showed that a purified PAL
protein interacted in vitro with pure peptidoglycan. A peptide competition experiment further demonstrated that the region from residues 89 to 130 of PAL was sufficient to bind the peptidoglycan. Moreover, the fact that this same region of PAL was also binding to
TolB suggested that these two interactions were exclusive. Indeed, the
TolB-PAL complex appeared not to be associated with the peptidoglycan.
This led us to the conclusion that PAL may exist in two forms in the
cell envelope, one bound to TolB and the other bound to the peptidoglycan.
*
Corresponding author. Present address: EMBL
Meyerhofstrasse-1, D-69117 Heidelberg, Germany. Phone: (49) (0) 6221 387492. Fax: (49) (0) 6221 387518. E-mail:
bouveret{at}embl-heidelberg.de.

Present address: Centre de Biophysique Moléculaire, 45071 Orleans Cedex 2,
France.
Journal of Bacteriology, October 1999, p. 6306-6311, Vol. 181, No. 20
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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