Functional reconstitution of beta-glucan elicitor-binding activity upon incorporation into lipid vesicles.

Abstract:

:In temperature-induced Triton X-114 phase separation experiments the beta-glucan elicitor-binding site from soybean (Glycine max L.) root membranes was identified as (a) hydrophobic membrane protein(s). The Zwittergent 3-12-solubilized beta-glucan-binding proteins were incorporated into lipid vesicles by the detergent-dilution procedure. Reconstituted binding proteins were functional in that binding of the hepta-beta-glucoside ligand was saturable, reversible and of high affinity (K(d)=6-7 nM). Competition studies using beta-glucans with different degrees of polymerization (DP 7-15; DP 15-25) showed effective displacement of the radioligand from the binding site whereas beta-glucan fragments with DP <7 were ineffective. The total amount of reconstituted binding activity was dependent on the acyl chain length of the phospholipids used for the reconstitution with a preference for decanoic (C10) and dodecanoic (C12) chains. Restored ligand binding was maximally 37% as compared to the former detergent-solubilized binding activity. The presence of a lipid environment stabilized the purified beta-glucan-binding proteins.

journal_name

FEBS Lett

journal_title

FEBS letters

authors

Mithöfer A,Ebel J

doi

10.1016/s0014-5793(99)01126-6

keywords:

subject

Has Abstract

pub_date

1999-09-17 00:00:00

pages

129-32

issue

2

eissn

0014-5793

issn

1873-3468

pii

S0014-5793(99)01126-6

journal_volume

458

pub_type

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