Characterization of the Staphylococcal enterotoxin A

Vβ receptor interaction using human receptor fragments engineered for high affinity

P. Sharma, S. Postel, E. J. Sundberg, David M Kranz

Research output: Contribution to journalArticle

Abstract

Staphylococcal food poisoning is a gastrointestinal disorder caused by the consumption of food containing Staphylococcal enterotoxins. Staphylococcal enterotoxin A (SEA) is the most common enterotoxin recovered from food poisoning outbreaks in the USA. In addition to its enteric activity, SEA also acts as a potent superantigen through stimulation of T cells, although less is known about its interactions than the superantigens SEB, SEC and toxic shock syndrome toxin-1. To understand more about SEA:receptor interactions, and to develop toxindetection systems for use in food testing, we engineered various SEA-binding receptor mutants. The extracellular domain of the receptor, a variable region of the beta chain (Vb22) of the T-cell receptor, was engineered for stability as a soluble protein and for high affinity, using yeastdisplay technology. The highest affinity mutant was shown to bind SEA with a Kd value of 4 nM. This was a 25 000- fold improvement in affinity compared with the wild-type receptor, which bound to SEA with low affinity (Kd value of 100 mM), similar to other superantigen:Vb interactions. The SEA:Vb interface was centered around residues within the complementarity determining region 2 loop. The engineered receptor was specific for SEA, in that it did not bind to two other closely related enterotoxins SEE or SED, providing information on the SEA residues possibly involved in the interaction. The specificity and affinity of these high-affinity Vb proteins also provide useful agents for the design of more sensitive and specific systems for SEA detection.

Original languageEnglish (US)
Pages (from-to)781-789
Number of pages9
JournalProtein Engineering, Design and Selection
Volume26
Issue number12
DOIs
StatePublished - Dec 1 2013

Fingerprint

T-cells
Enterotoxins
Proteins
Superantigens
Staphylococcal Food Poisoning
Staphylococcal enterotoxin A
Complementarity Determining Regions
Testing
Antigen Receptors, T-Cell, alpha-beta
Food
Foodborne Diseases
Disease Outbreaks
Technology
T-Lymphocytes

Keywords

  • Directed evolution
  • Food poisoning
  • Staphylococcal enterotoxin A (SEA)
  • Yeast display

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Molecular Biology

Cite this

Characterization of the Staphylococcal enterotoxin A : Vβ receptor interaction using human receptor fragments engineered for high affinity. / Sharma, P.; Postel, S.; Sundberg, E. J.; Kranz, David M.

In: Protein Engineering, Design and Selection, Vol. 26, No. 12, 01.12.2013, p. 781-789.

Research output: Contribution to journalArticle

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