Defining the Structural Basis of Human Plasminogen Binding by Streptococcal Surface Enolase

Overview

Journal J Biol Chem

Specialty Biochemistry

Date 2009 Apr 14

PMID 19363026

Citations 38

Authors

Amanda J Cork

Slobodan Jergic

Sven Hammerschmidt

Bostjan Kobe

Vijay Pancholi

Justin L P Benesch

Carol V Robinson

Nicholas E Dixon

J Andrew Aquilina

Mark J Walker

Affiliations

Soon will be listed here.

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 alpha-enolase. Site-directed mutagenesis of surface-located lysine residues (SEN(K252 + 255A), SEN(K304A), SEN(K334A), SEN(K344E), SEN(K435L), and SEN(Delta434-435)) 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 SEN(K344E), 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 SEN(K344E) had lost alpha-enolase activity, which was also reduced in SEN(K334A) and SEN(Delta434-435). Surface plasmon resonance demonstrated that the capacity to bind human plasminogen was abolished in SEN(K252 + 255A), SEN(K435L), and SEN(Delta434-435). 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.

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References

Benesch J . Collisional activation of protein complexes: picking up the pieces. J Am Soc Mass Spectrom. 2008; 20(3):341-8. DOI: 10.1016/j.jasms.2008.11.014. View

Cole J, Ramirez R, Currie B, Cordwell S, Djordjevic S, Walker M . Surface analyses and immune reactivities of major cell wall-associated proteins of group a streptococcus. Infect Immun. 2005; 73(5):3137-46. PMC: 1087385. DOI: 10.1128/IAI.73.5.3137-3146.2005. View

Carapetis J, Walker A, Hibble M, Sriprakash K, Currie B . Clinical and epidemiological features of group A streptococcal bacteraemia in a region with hyperendemic superficial streptococcal infection. Epidemiol Infect. 1999; 122(1):59-65. PMC: 2809588. DOI: 10.1017/s0950268898001952. View

Sanderson-Smith M, Batzloff M, Sriprakash K, Dowton M, Ranson M, Walker M . Divergence in the plasminogen-binding group a streptococcal M protein family: functional conservation of binding site and potential role for immune selection of variants. J Biol Chem. 2005; 281(6):3217-26. DOI: 10.1074/jbc.M508758200. View

Benesch J, Aquilina J, Ruotolo B, Sobott F, Robinson C . Tandem mass spectrometry reveals the quaternary organization of macromolecular assemblies. Chem Biol. 2006; 13(6):597-605. DOI: 10.1016/j.chembiol.2006.04.006. View

Eswar N, John B, Mirkovic N, Fiser A, Ilyin V, Pieper U . Tools for comparative protein structure modeling and analysis. Nucleic Acids Res. 2003; 31(13):3375-80. PMC: 168950. DOI: 10.1093/nar/gkg543. View

Sanderson-Smith M, Dowton M, Ranson M, Walker M . The plasminogen-binding group A streptococcal M protein-related protein Prp binds plasminogen via arginine and histidine residues. J Bacteriol. 2006; 189(4):1435-40. PMC: 1797364. DOI: 10.1128/JB.01218-06. View

Parry M, Zhang X, Bode I . Molecular mechanisms of plasminogen activation: bacterial cofactors provide clues. Trends Biochem Sci. 2000; 25(2):53-9. DOI: 10.1016/s0968-0004(99)01521-2. View

Tart A, Walker M, Musser J . New understanding of the group A Streptococcus pathogenesis cycle. Trends Microbiol. 2007; 15(7):318-25. DOI: 10.1016/j.tim.2007.05.001. View

10.

Sreerama N, Woody R . Estimation of protein secondary structure from circular dichroism spectra: comparison of CONTIN, SELCON, and CDSSTR methods with an expanded reference set. Anal Biochem. 2000; 287(2):252-60. DOI: 10.1006/abio.2000.4880. View

11.

Johnson M, Zaretskaya I, Raytselis Y, Merezhuk Y, McGinnis S, Madden T . NCBI BLAST: a better web interface. Nucleic Acids Res. 2008; 36(Web Server issue):W5-9. PMC: 2447716. DOI: 10.1093/nar/gkn201. View

12.

Carapetis J, Steer A, Mulholland E, Weber M . The global burden of group A streptococcal diseases. Lancet Infect Dis. 2005; 5(11):685-94. DOI: 10.1016/S1473-3099(05)70267-X. View

13.

Kuusela P, Ullberg M, Saksela O, Kronvall G . Tissue-type plasminogen activator-mediated activation of plasminogen on the surface of group A, C, and G streptococci. Infect Immun. 1992; 60(1):196-201. PMC: 257522. DOI: 10.1128/iai.60.1.196-201.1992. View

14.

Riekel C, Burghammer M, Schertler G . Protein crystallography microdiffraction. Curr Opin Struct Biol. 2005; 15(5):556-62. DOI: 10.1016/j.sbi.2005.08.013. View

15.

Byrne B, Iwata S . Membrane protein complexes. Curr Opin Struct Biol. 2002; 12(2):239-43. DOI: 10.1016/s0959-440x(02)00316-0. View

16.

Benesch J, Robinson C . Mass spectrometry of macromolecular assemblies: preservation and dissociation. Curr Opin Struct Biol. 2006; 16(2):245-51. DOI: 10.1016/j.sbi.2006.03.009. View

17.

Wang H, Lottenberg R, Boyle M . A role for fibrinogen in the streptokinase-dependent acquisition of plasmin(ogen) by group A streptococci. J Infect Dis. 1995; 171(1):85-92. DOI: 10.1093/infdis/171.1.85. View

18.

McKay F, McArthur J, Sanderson-Smith M, Gardam S, Currie B, Sriprakash K . Plasminogen binding by group A streptococcal isolates from a region of hyperendemicity for streptococcal skin infection and a high incidence of invasive infection. Infect Immun. 2003; 72(1):364-70. PMC: 343955. DOI: 10.1128/IAI.72.1.364-370.2004. View

19.

Walker M, Hollands A, Sanderson-Smith M, Cole J, Kirk J, Henningham A . DNase Sda1 provides selection pressure for a switch to invasive group A streptococcal infection. Nat Med. 2007; 13(8):981-5. DOI: 10.1038/nm1612. View

20.

Jordan P, Fromme P, Witt H, Klukas O, Saenger W, Krauss N . Three-dimensional structure of cyanobacterial photosystem I at 2.5 A resolution. Nature. 2001; 411(6840):909-17. DOI: 10.1038/35082000. View