[en] Highlights: • An NMR assignment of Lam4S2 identified significant conformational exchange. • Lam4S2 binding to sterol confirms the StART-like cavity as its binding site. • Using MβCD as cholesterol donor/acceptor, the upper limit of sterol exchange is ∼6 s⁻¹. Sterols are essential components of cellular membranes and shape their biophysical properties. The recently discovered family of Lipid transfer proteins Anchored at Membrane contact sites (LAMs) has been suggested to carry out intracellular sterol traffic using StART-like domains. Here, we studied the second StART-like domain of Lam4p from S. cerevisiae by NMR. We show that NMR data are consistent with the StART-like domain structure, and that several functionally important regions within the domain exhibit significant conformational dynamics. NMR titration experiments confirm sterol binding to the canonical sterol-binding site and suggest a role of membrane interactions on the thermodynamics and kinetics of sterol binding.