HrpZPsph interacts with lipid membranes. (A)
Psph race-6 wild-type (wt) and race-6 mutant hrpA− were
grown in hrp-inducing minimal media (8) in the absence
(−) or presence (+) of liposomes. Proteins prepared from the
supernatant or proteoliposomes were separated and analyzed by
SDS/PAGE immunoblotting with an anti-HrpZPsph antiserum.
(B) TRANSIL beads coated with
1-hexadecanoyl-2-(cis-9-octadecenoyl)-sn-glycero-3-phosphocholine
(POPC)/1-hexadecanoyl-2-(cis-9-octadecenoyl)-sn-glycero-3-phosphoethanolamine
(POPE) (80:20) were incubated for 1 h with 1 μM of the proteins
indicated (Total). After separation of lipid-bound (Bound) from unbound
material, proteins were analyzed by SDS/PAGE and silver staining.
HrpZPst, HrpZ from P. syringae pv.
tomato DC3000; HrpZPss, HrpZ from P.
syringae pv. syringae; AA 1–80, AA 100–200, AA
201–345, HrpZPsph fragment encompassing the N-terminal 80
amino acid (AA) residues, amino acids 100–200, or the C-terminal amino
acids 201–345, respectively.