Journal of Biomolecular NMR

, Volume 40, Issue 3, pp 213–217 | Cite as

NMR structure note: alkaline proteinase inhibitor APRin from Pseudomonas aeruginosa

  • Sengodagounder Arumugam
  • Robert D. Gray
  • Andrew N. Lane
NMR Structure Note

Biological context

The alkaline proteinase inhibitor (APRin) from Pseudomonas aeruginosa is an 11.5-kDa, high affinity, high specificity inhibitor of the serralysin class of zinc-dependent proteinases secreted by several Gram-negative bacteria (Feltzer et al. 2000). The inhibitor APRin is co-secreted from the bacterium presumably to prevent adventitious proteolysis of host proteins during the secretion process. The serralysins are mechanistically and structurally related to the matrix metalloproteinases (Morihara et al. 2000), and the active site zinc atom is accessible through a tunnel to the enzyme surface. These enzymes are capable of degrading a variety of host proteins and thereby enhance the pathogenicity of these organisms (Morihara and Homma 1985).

The X-ray structure of the proteinase-APRin complex revealed that the five N-terminal inhibitor residues occupy the extended substrate binding site of the enzyme and that the terminal amino group (Ser) coordinates to the catalytic...


Extended Chain Rotational Correlation Time Residual Dipolar Coupling Catalytic Zinc Backbone Chemical Shift 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



This work was supported by the Kentucky Challenge for Excellence (to ANL). NMR spectra were recorded at the JG Brown Cancer Center NMR Facility with support from The National Science Foundation EPSCoR grant # EPS−0447479 and the Brown Foundation.

Supplementary material

10858_2008_9218_MOESM1_ESM.doc (1.8 mb)
(DOC 1810 kb)


  1. Arumugam S, Gray RD, Lane AN (2005) H-1, N-15 and C-13 assignments of the alkaline proteinase inhibitor APRin from Pseudomonas aeruginosa. J Biomol NMR 31:265–266CrossRefGoogle Scholar
  2. Cavanagh J, Fairbrother WJ, Palmer AG, Skelton AGNJ (1996) Protein NMR spectroscopy principles and practice. Academic Press, San DiegoGoogle Scholar
  3. Cornilescu G, Delaglio F, Bax A (1999) Protein backbone angle restraints from searching a database for chemical shift and sequence homology. J Biomol NMR 13:289–302CrossRefGoogle Scholar
  4. Delaglio F, Grzesiek S, Vuister GW, Zhu G, Pfeifer J, Bax A (1995) Nmrpipe—a multidimensional spectral processing system based on unix pipes. J Biomol NMR 6:277–293CrossRefGoogle Scholar
  5. Dyson HJ, Wright PE (2005) Intrinsically unstructured proteins and their functions. Nature Rev Mol Cell Biol 6:197–208CrossRefGoogle Scholar
  6. Feltzer RE, Gray RD, Dean WL, Pierce WM (2000) Alkaline proteinase inhibitor of Pseudomonas aeruginosa—interaction of native and N-terminally truncated inhibitor proteins with Pseudomonas metalloproteinases. J Biol Chem 275:21002–21009CrossRefGoogle Scholar
  7. Feltzer RE, Trent JO, Gray RD (2003) Alkaline proteinase inhibitor of Pseudomonas aeruginosa—a mutational and molecular dynamics study of the role of N-terminal residues in the inhibition of Pseudomonas alkaline proteinase. J Biol Chem 278:25952–25957CrossRefGoogle Scholar
  8. Gray RD, Trent JO (2005) Contribution of a single-turn alpha-helix to the conformational stability and activity of the alkaline proteinase inhibitor of Pseudomonas aeruginosa. Biochemistry 44:2469–2477CrossRefGoogle Scholar
  9. Hall JB, Fushman D (2006) Variability of the N-15 chemical shielding tensors in the B3 domain of protein G from N-15 relaxation measurements at several fields. Implications for backbone order parameters. J Am Chem Soc 128:7855–7870CrossRefGoogle Scholar
  10. Hege T, Feltzer RE, Gray RD, Baumann U (2001) Crystal structure of a complex between Pseudomonas aeruginosa alkaline protease and its cognate inhibitor—inhibition by a zinc-NH2 coordinative bond. J Biol Chem 276:35087–35092CrossRefGoogle Scholar
  11. Kay LE, Nicholson LK, Delaglio F, Bax A, Torchia DA (1992) Pulse sequences for removal of the effects of cross-correlation between dipolar and chemical-shift anisotropy relaxation mechanism on the measurement of heteronuclear T1 and T2 values in proteins. J Magn Reson 97:359–375Google Scholar
  12. Laskowski RA, Rullmann JAC, MacArthur MW, Kaptein R, Thornton JM (1996) AQUA and PROCHECK-NMR: programs for checking the quality of protein structures solved by NMR. J Biomol NMR 8:477–486CrossRefGoogle Scholar
  13. Marley J, Lu M, Bracken C (2001) A method for efficient isotopic labeling of recombinant proteins. J Biomol NMR 20:71–75CrossRefGoogle Scholar
  14. McIntosh PB, Taylor IA, Frenkiel TA, Smerdon SJ, Lane AN (2000) The influence of DNA binding on the backbone dynamics of the yeast cell-cycle protein Mbp1. J Biomol NMR 16:183–196CrossRefGoogle Scholar
  15. Morihara K, Homma J (1985) Bacterial enzymes and virulence bacterial enzymes and virulence I. Holder. CRC Press, Boca Raton, pp 41–47Google Scholar
  16. Morihara K, Hata Y, Okuda K (2000) Serralysin Zn-metalloproteinases—structure, function, secretion pathway, and pathogenicity. Seikagaku 72:16–25Google Scholar
  17. Piotto M, Saudek V, Sklenar V (1992) Gradient-tailored excitation for single-quantum NMR spectroscopy of aqueous solutions. J Biomol NMR 2:661–665CrossRefGoogle Scholar
  18. Ruckert M, Otting G (2000) Alignment of biological macromolecules in novel nonionic liquid crystalline media for NMR experiments. J Am Chem Soc 122:7793–7797CrossRefGoogle Scholar
  19. Schwieters CD, Kuszewski JJ, Clore GM (2006) Using Xplor-NIH for NMR molecular structure determination. Prog NMR Spectrosc 48:47–62CrossRefGoogle Scholar
  20. Wishart DS, Bigam CG, Holm A, Hodges RS, Sykes BD (1995) H-1, C-13 and N-15 random coil NMR chemical-shifts of the common amino-acids .1. Investigations of nearest-neighbor effects. J Biomol NMR 5:67–81CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • Sengodagounder Arumugam
    • 1
  • Robert D. Gray
    • 2
  • Andrew N. Lane
    • 1
  1. 1.J.G. Graham Brown Cancer Center, University of Louisville LouisvilleUSA
  2. 2.Department of Biochemistry and Molecular BiologyUniversity of LouisvilleLouisvilleUSA

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