The structural characteristics of nonspecific lipid transfer proteins explain their resistance to gastroduodenal proteolysis

Ramani Wijesinha-Bettoni, Yuri Alexeev, Phil Johnson, Justin Marsh, Ana I. Sancho, Syed U. Abdullah, Alan R. Mackie, Peter R. Shewry, Lorna J. Smith, E. N. Clare Mills

Research output: Contribution to journalArticlepeer-review

39 Scopus citations


The structure and stability of the allergenic nonspecific lipid transfer protein (LTP) of peach were compared with the homologous LTP1 of barley and its liganded form LTP1b. All three proteins were resistant to gastric pepsinolysis and were only slowly digested at 1 to 2 out of 14 potential tryptic and chymotryptic cleavage sites under duodenal conditions. Peach LTP was initially cleaved at Tyr79-Lys80 and then at Arg39-Thr40 (a site lost in barley LTP1). Molecular dynamics simulations of the proteins under folded conditions showed that the backbone flexibility is limited, explaining the resistance to duodenal proteolysis. Arg39 and Lys80 side chains were more flexible in simulations of peach, compared with, barley LTP1. This may explain differences in the rates of cleavage observed experimentally for the two proteins and suggests that the flexibility of individual amino acid side chains could be important in determining preferred proteolytic cleavage sites. In order to understand resistance to pepsinolysis, proteins were characterized by NMR spectroscopy at pH 1.8. This showed that the helical regions of both proteins remain folded at this pH. NMR hydrogen exchange studies confirmed the rigidity of the structures at acidic pH, with barley LTP1 showing some regions with greater protection. Collectively, these data suggest that the rigidity of the LTP scaffold is responsible for their resistance to proteolysis. Gastroduodenal digestion conditions do not disrupt the 3D structure of peach LTP, explaining why LTPs retain their ability to bind IgE after digestion and hence their allergenic potential.

Original languageEnglish (US)
Pages (from-to)2130-2139
Number of pages10
Issue number10
StatePublished - Mar 16 2010
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry


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