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Biochemistry. 2007 Dec 4;46(48):13929-37. Epub 2007 Nov 7.

Puroindoline-b mutations control the lipid binding interactions in mixed puroindoline-a:puroindoline-b systems.

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1
Department of Food Biosciences, University of Reading, PO Box 226, Whiteknights, Reading RG6 6AP, U.K.

Abstract

The interactions have been investigated of puroindoline-a (Pin-a) and mixed protein systems of Pin-a and wild-type puroindoline-b (Pin-b+) or puroindoline-b mutants (G46S mutation (Pin-bH) or W44R mutation (Pin-bS)) with condensed phase monolayers of an anionic phospholipid (L-alpha-dipalmitoylphosphatidyl-dl-glycerol (DPPG)) at the air/water interface. The interactions of the mixed systems were studied at three different concentration ratios of Pin-a:Pin-b, namely 3:1, 1:1 and 1:3 in order to establish any synergism in relation to lipid binding properties. Surface pressure measurements revealed that Pin-a interaction with DPPG monolayers led to an equilibrium surface pressure increase of 8.7 +/- 0.6 mN m-1. This was less than was measured for Pin-a:Pin-b+ (9.6 to 13.4 mN m-1), but was significantly more than was measured for Pin-a:Pin-bH (4.0 to 6.2 mN m-1) or Pin-a:Pin-bS (3.8 to 6.3 mN m-1) over the complete range of concentration ratio. Consequently, surface pressure increases were shown to correlate to endosperm hardness phenotype, with puroindolines present in hard-textured wheat varieties yielding lower equilibrium surface pressure changes. Integrated amide I peak areas from corresponding external reflectance Fourier-transform infrared (ER-FTIR) spectra, used to indicate levels of protein adsorption to the lipid monolayers, showed that differences in adsorbed amount were less significant. The data therefore suggest that Pin-b mutants having single residue substitutions within their tryptophan-rich loop that are expressed in some hard-textured wheat varieties influence the degree of penetration of Pin-a and Pin-b into anionic phospholipid films. These findings highlight the key role of the tryptophan-rich loop in puroindoline-lipid interactions.

PMID:
17985936
DOI:
10.1021/bi701680w
[Indexed for MEDLINE]
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