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PMID: 12324323 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Replacement of trifluoroacetic acid with HCl in the hydrophobic purification steps of pediocin PA-1: a structural effect.

Applied and environmental microbiology ·Vol. 68 ·No. 10 ·2002-10-00 ·Pages 4803-8

Gaussier H, Morency H, Lavoie MC, Subirade M

Abstract

Trifluoroacetic acid (TFA) is a purification contaminant associated with pediocin PA-1 that interferes with Fourier transform infrared spectroscopy structural analysis. As revealed by circular dichroism, its presence affects the structural folding of pediocin. Consequently, we propose a new pediocin PA-1 purification procedure using HCl instead of TFA in all of the hydrophobic steps. This procedural change does not affect the purification yield or the amount of pediocin PA-1 purified. Furthermore, removing HCl, as opposed to TFA, after purification is an easier procedure to carry out. In fact, the removal of TFA requires more experimentation and results in protein loss. Thus, HCl is a good alternative to TFA in pediocin PA-1 purification and can be extended to the purification of other proteins. We also show that TFA-induced structural modifications do not significantly affect the antimicrobial activity of pediocin PA-1.

MeSH Terms
Bacteriocins/chemistry,isolation & purification Circular Dichroism Hydrochloric Acid/chemistry Mass Spectrometry Pediocins Spectrophotometry, Ultraviolet Trifluoroacetic Acid/chemistry
Chemicals
Bacteriocins Pediocins pediocin PA-1 Trifluoroacetic Acid Hydrochloric Acid
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Gaussier Hélène
Centre de Recherche en Sciences et Technologie du Lait, Chaire de Recherche du Canada sur les Protéines et les Aliments Fonctionnels, and Département des Sciences des Aliments et de Nutrition, Université Laval, Sainte-Foy, Québec G1K 7P4, Canada.
Morency Hélène
Lavoie Marc C
Subirade Muriel
References (39)
39 references, click to expand
  1. Fourier transform infrared spectroscopic studies of the interaction of the antimicrobial peptide gramicidin S with lipid micelles and with lipid monolayer and bilayer membranes.
    Biochemistry. 1999 Nov 16;38(46):15193-203 PMID: 10563802
  2. Androctonin, a hydrophilic disulphide-bridged non-haemolytic anti-microbial peptide: a plausible mode of action.
    Biochem J. 2000 Feb 1;345 Pt 3:653-64 PMID: 10642525
  3. Trifluoroacetate, a contaminant in purified proteins, inhibits proliferation of osteoblasts and chondrocytes.
    Am J Physiol. 1999 Nov;277(5 Pt 1):E779-83 PMID: 10567002
  4. Solution structure of carnobacteriocin B2 and implications for structure-activity relationships among type IIa bacteriocins from lactic acid bacteria.
    Biochemistry. 1999 Nov 23;38(47):15438-47 PMID: 10569926
  5. Method for rapid purification of class IIa bacteriocins and comparison of their activities.
    Appl Environ Microbiol. 2000 Apr;66(4):1744-8 PMID: 10742275
  6. Site-specific conformational determination in thermal unfolding studies of helical peptides using vibrational circular dichroism with isotopic substitution.
    Proc Natl Acad Sci U S A. 2000 Jul 18;97(15):8318-23 PMID: 10880566
  7. Trichloroacetic acid and trifluoroacetic acid-induced unfolding of cytochrome c: stabilization of a native-like folded intermediate(1).
    Biochim Biophys Acta. 2000 Jul 14;1480(1-2):201-10 PMID: 11004564
  8. Spontaneous insertion of gene 9 minor coat protein of bacteriophage M13 in model membranes.
    Biochim Biophys Acta. 2001 Apr 2;1511(2):309-16 PMID: 11286974
  9. Conformational rearrangement of beta-lactoglobulin upon interaction with an anionic membrane.
    Biochim Biophys Acta. 2001 Sep 10;1549(1):37-50 PMID: 11566367
  10. Conformational changes in pediocin AcH upon vesicle binding and approximation of the membrane-bound structure in detergent micelles.
    Biochemistry. 2001 Nov 20;40(46):14037-46 PMID: 11705396
  11. Bacteriocins of gram-positive bacteria.
    Bacteriol Rev. 1976 Sep;40(3):722-56 PMID: 791239
  12. Conformation of DNA in chromatin reconstituted from poly [d(A-T)] and the core histones.
    Nucleic Acids Res. 1981 Oct 10;9(19):4879-93 PMID: 7312620
  13. New insight into protein secondary structure from resolution-enhanced infrared spectra.
    Biochim Biophys Acta. 1988 Jan 29;952(2):115-30 PMID: 3276352
  14. Secondary structure of proteins through circular dichroism spectroscopy.
    Annu Rev Biophys Biophys Chem. 1988;17:145-66 PMID: 3293583
  15. Bacteriocins of lactic acid bacteria.
    Biochimie. 1988 Mar;70(3):337-49 PMID: 3139051
  16. Protein chemistry of the Neurospora crassa plasma membrane H+-ATPase.
    Anal Biochem. 1988 Sep;173(2):251-64 PMID: 2903697
  17. Inhibition of Listeria monocytogenes by using bacteriocin PA-1 produced by Pediococcus acidilactici PAC 1.0.
    Appl Environ Microbiol. 1988 Oct;54(10):2349-53 PMID: 3144243
  18. Purification, characterization and antimicrobial spectrum of a bacteriocin produced by Pediococcus acidilactici.
    J Appl Bacteriol. 1988 Oct;65(4):261-8 PMID: 2906056
  19. Preliminary characterization of four bacteriocins from Streptococcus mutans.
    Can J Microbiol. 1990 Feb;36(2):123-30 PMID: 2337834
  20. Amide modes and protein conformation.
    Biochim Biophys Acta. 1992 Apr 8;1120(2):123-43 PMID: 1373323
  21. Purification and primary structure of pediocin PA-1 produced by Pediococcus acidilactici PAC-1.0.
    Arch Biochem Biophys. 1992 May 15;295(1):5-12 PMID: 1575516
  22. Conformation of magainin-2 and related peptides in aqueous solution and membrane environments probed by Fourier transform infrared spectroscopy.
    Biochemistry. 1992 Aug 18;31(32):7289-93 PMID: 1510920
  23. Novel method to extract large amounts of bacteriocins from lactic acid bacteria.
    Appl Environ Microbiol. 1992 Oct;58(10):3355-9 PMID: 1444369
  24. Quantitative studies of the structure of proteins in solution by Fourier-transform infrared spectroscopy.
    Prog Biophys Mol Biol. 1993;59(1):23-56 PMID: 8419985
  25. Evaluation of secondary structure of proteins from UV circular dichroism spectra using an unsupervised learning neural network.
    Protein Eng. 1993 Jun;6(4):383-90 PMID: 8332596
  26. On the biosynthesis of bovine pancreatic trypsin inhibitor (BPTI). Structure, processing, folding and disulphide bond formation of the precursor in vitro and in microsomes.
    J Mol Biol. 1993 Aug 20;232(4):1176-96 PMID: 7690407
  27. Genetics of bacteriocins produced by lactic acid bacteria.
    FEMS Microbiol Rev. 1993 Sep;12(1-3):39-85 PMID: 8398217
  28. Determination of soluble and membrane protein structure by Fourier transform infrared spectroscopy. I. Assignments and model compounds.
    Subcell Biochem. 1994;23:329-62 PMID: 7855877
  29. The use and misuse of FTIR spectroscopy in the determination of protein structure.
    Crit Rev Biochem Mol Biol. 1995;30(2):95-120 PMID: 7656562
  30. Maturation pathway of nisin and other lantibiotics: post-translationally modified antimicrobial peptides exported by gram-positive bacteria.
    Mol Microbiol. 1995 Aug;17(3):427-37 PMID: 8559062
  31. Covalent structure, synthesis, and structure-function studies of mesentericin Y 105(37), a defensive peptide from gram-positive bacteria Leuconostoc mesenteroides.
    J Biol Chem. 1996 Jun 14;271(24):14421-9 PMID: 8662868
  32. The different molar absorptivities of the secondary structure types in the amide I region: an attenuated total reflection infrared study on globular proteins.
    Anal Biochem. 1996 Nov 1;242(1):95-103 PMID: 8923971
  33. Purification and amino acid sequences of piscicocins V1a and V1b, two class IIa bacteriocins secreted by Carnobacterium piscicola V1 that display significantly different levels of specific inhibitory activity.
    Appl Environ Microbiol. 1996 Dec;62(12):4410-6 PMID: 8953713
  34. Purification and structure of mutacin B-Ny266: a new lantibiotic produced by Streptococcus mutans.
    FEBS Lett. 1997 Jun 30;410(2-3):275-9 PMID: 9237644
  35. Three-dimensional structure of leucocin A in trifluoroethanol and dodecylphosphocholine micelles: spatial location of residues critical for biological activity in type IIa bacteriocins from lactic acid bacteria.
    Biochemistry. 1997 Dec 9;36(49):15062-72 PMID: 9398233
  36. Secondary structure analysis of the putative membrane-associated domains of the inward rectifier K+ channel ROMK1.
    Biochem J. 1998 Oct 15;335 ( Pt 2):375-80 PMID: 9761737
  37. Generation of polyclonal antibodies of predetermined specificity against pediocin PA-1.
    Appl Environ Microbiol. 1998 Nov;64(11):4536-45 PMID: 9797320
  38. Divercin V41, a new bacteriocin with two disulphide bonds produced by Carnobacterium divergens V41: primary structure and genomic organization.
    Microbiology. 1998 Oct;144 ( Pt 10):2837-44 PMID: 9802025
  39. 2D-NMR and ATR-FTIR study of the structure of a cell-selective diastereomer of melittin and its orientation in phospholipids.
    Biochemistry. 1999 Nov 16;38(46):15305-16 PMID: 10563816
Article Info
Journal
Applied and environmental microbiology
Abbr.
Appl Environ Microbiol
ISSN
0099-2240
DOI
10.1128/AEM.68.10.4803-4808.2002
Published
2002-10-00
Pages
4803-8
Language
English
Region
United States
NLM ID
7605801
PMCID
PMC126396
Subset
IM
External Links
PubMed source DOI: 10.1128/AEM.68.10.4803-4808.2002 PMC Full Text View journal details
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