Defining the structural basis of human plasminogen binding by streptococcal surface enolase

Cork, A. J., Jergic, S., Hammerschmidt, S., Kobe, B., Pancholi, V., Benesch, J. L. P., Robinson, C. V., Dixon, N. E., Aquilina, J. A. and Walker, M. J. (2009) Defining the structural basis of human plasminogen binding by streptococcal surface enolase. Journal of Biological Chemistry, 284 25: 17129-17137. doi:10.1074/jbc.M109.004317

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Author Cork, A. J.
Jergic, S.
Hammerschmidt, S.
Kobe, B.
Pancholi, V.
Benesch, J. L. P.
Robinson, C. V.
Dixon, N. E.
Aquilina, J. A.
Walker, M. J.
Title Defining the structural basis of human plasminogen binding by streptococcal surface enolase
Journal name Journal of Biological Chemistry   Check publisher's open access policy
ISSN 0021-9258
Publication date 2009-06-19
Sub-type Article (original research)
DOI 10.1074/jbc.M109.004317
Open Access Status File (Publisher version)
Volume 284
Issue 25
Start page 17129
End page 17137
Total pages 9
Place of publication Bethesda, MD, U.S.A.
Publisher American Society for Biochemistry and Molecular Biology
Collection year 2010
Language eng
Subject C1
060109 Proteomics and Intermolecular Interactions (excl. Medical Proteomics)
970106 Expanding Knowledge in the Biological Sciences
Formatted abstract
The flesh-eating bacterium group A Streptococcus (GAS) binds and activates human plasminogen, promoting invasive disease. Streptococcal surface enolase (SEN), a glycolytic pathway enzyme, is an identified plasminogen receptor of GAS. Here we used mass spectrometry (MS) to confirm that GAS SEN is octameric, thereby validating in silico modeling based on the crystal structure of Streptococcus pneumoniae α-enolase. Site-directed mutagenesis of surface-located lysine residues (SENK252 + 255A, SENK304A, SENK334A, SENK344E, SENK435L, and SENΔ434–435) was used to examine their roles in maintaining structural integrity, enzymatic function, and plasminogen binding. Structural integrity of the GAS SEN octamer was retained for all mutants except SENK344E, as determined by circular dichroism spectroscopy and MS. However, ion mobility MS revealed distinct differences in the stability of several mutant octamers in comparison with wild type. Enzymatic analysis indicated that SENK344E had lost α-enolase activity, which was also reduced in SENK334A and SENΔ434–435. Surface plasmon resonance demonstrated that the capacity to bind human plasminogen was abolished in SENK252 + 255A, SENK435L, and SENΔ434–435. The lysine residues at positions 252, 255, 434, and 435 therefore play a concerted role in plasminogen acquisition. This study demonstrates the ability of combining in silico structural modeling with ion mobility-MS validation for undertaking functional studies on complex protein structures.
© 2009 by The American Society for Biochemistry and Molecular Biology, Inc.
Keyword Group-A streptococci
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

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Created: Thu, 03 Sep 2009, 08:00:03 EST by Mr Andrew Martlew on behalf of School of Chemistry & Molecular Biosciences