Detection and identification of virulence factors in Yersinia pestis using SELDI ProteinChip® system

V. Thulasiraman, S. L. McCutchen-Maloney, Vladimir Motin, E. Garcia

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

A rapid method for the detection, purification, and identification of proteins in bacterial extracts was developed using surface enhanced laser desorption/ionization (SELDI) ProteinChip® technology. The effectiveness of this technique for monitoring the expression and identification of temperature- and calcium-regulated virulence factors of Yersinia pestis, the bacterium that causes human plague, is demonstrated. Y. pestis infection of its mammalian host is thought to be accompanied by rapid up-regulation of a number of genes following a shift from 26°C (the temperature of the flea vector) to 37°C (the temperature of the mammalian host). To model this process, Y. pestis cells were grown at 26°C and 37°C in a Ca2+-deficient medium. Through an initial protein profiling of the crude bacterial extract on strong anion exchange and copper affinity, ProteinChip arrays detected five proteins that were up-regulated and three proteins that were down-regulated at 37°C. Two of the proteins predominately expressed at 37°C were semi-purified in less than two days. The two proteins were identified as catalase-peroxidase and Antigen 4. Aside from its speed, a salient feature of the SELDI technique is the microgram amounts of crude sample required for analysis.

Original languageEnglish (US)
Pages (from-to)428-432
Number of pages5
JournalBioTechniques
Volume30
Issue number2
StatePublished - 2001
Externally publishedYes

Fingerprint

Yersinia pestis
Protein Array Analysis
Virulence Factors
Ionization
Desorption
Lasers
Proteins
Temperature
Yersinia Infections
Siphonaptera
Bacterial Proteins
Plague
Complex Mixtures
Catalase
Peroxidase
Anions
Copper
Up-Regulation
Purification
Bacteria

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Thulasiraman, V., McCutchen-Maloney, S. L., Motin, V., & Garcia, E. (2001). Detection and identification of virulence factors in Yersinia pestis using SELDI ProteinChip® system. BioTechniques, 30(2), 428-432.

Detection and identification of virulence factors in Yersinia pestis using SELDI ProteinChip® system. / Thulasiraman, V.; McCutchen-Maloney, S. L.; Motin, Vladimir; Garcia, E.

In: BioTechniques, Vol. 30, No. 2, 2001, p. 428-432.

Research output: Contribution to journalArticle

Thulasiraman, V, McCutchen-Maloney, SL, Motin, V & Garcia, E 2001, 'Detection and identification of virulence factors in Yersinia pestis using SELDI ProteinChip® system', BioTechniques, vol. 30, no. 2, pp. 428-432.
Thulasiraman, V. ; McCutchen-Maloney, S. L. ; Motin, Vladimir ; Garcia, E. / Detection and identification of virulence factors in Yersinia pestis using SELDI ProteinChip® system. In: BioTechniques. 2001 ; Vol. 30, No. 2. pp. 428-432.
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