Fig. S5.

Particular regions of Lam6p and Tco89p directly interact with tVac8p. (A) Schematic representations showing domain structures of Lam6p and Tco89p. A series of truncated constructs used for pull-down experiments are indicated. All constructs were fused to the His₆-SMT3 tag at their N termini. (B) His-tag pull-down experiments showing tVac8p interactions with Lam6p and Tco89p. GST-tagged tVac8p was coexpressed with His₆-SMT3-tagged Lam6p or Tco89p and immobilized on Ni²⁺-NTA resin. Experiments were carried out as described in Fig. 4B. (C) To test whether Vac8p shares the binding interface for Nvj1p with Lam6p or Tco89p, wild-type and triple mutant (R276E/R317E/R359E) GST-tagged tVac8p were coexpressed with his-tagged Lam6p (residues 400–628) or Tco89p (residues 401–600) and immobilized on Ni²⁺-NTA resin. Quantitative data are shown on the right. TM, transmembrane domain.