Conformational studies on an isolated integral membrane protein are reported. Lipoprotein of the Escherichia coli outer membrane was released from murein by treatment with either lysozyme or trypsin. The isolated lysozyme-released lipoprotein (lipoprotein I) contained 2 or 3 muropeptides covalently linked at the C-terminal end, while the trypsin-released lipoprotein (lipoprotein IT) was free of muropeptides and lacked the C-terminal peptide Tyr-Arg-Lys. Circular dichroism spectra of the two preparations were essentially identical, and they show an a-helix content of about 80%. According to calculations based on the Chou-Fasman rules for proteins of known sequence, lipoprotein is 64 % a-helix and 15 % p-structure. Infrared spectroscopy qualitatively supports these values. The conformation was stable in the pH range of 5-12. Denaturation of lipoprotein by heat, 8 M urea, or sodium dodecylsulphate was a fully reversible, cooperative process. The thermal denaturation of lipoprotein occurs in two steps with transition points at 79.4 "C for lipoprotein I and at 85.1 "C for lipoprotein 11. Lipoprotein markedly changes conformation at dodecylsulphate concentrations where micelle formation sets in. The unusual behaviour of the lipoprotein conformation in sodium dodecylsulphate is discussed in relation to the lipoprotein conformation and aggregation within the membrane.The conformation of membrane proteins has played an important role in discussions on the molecular organization of biological membranes [I -41. The outer membrane of Escherichia coli is basically constructed like other biological membranes in that glycolipids are at the surface, and phospholipids and proteins are the major constituents. A peculiarity of the bacterial cell's outer layer is the murein net composed of polysaccharide chains cross-linked by short peptide side-chains [5]. Lipoprotein molecules of a single type are covalently fixed at the murein net and extend into the outer membrane [6,7]. Together with the free form of lipoprotein not covalently bound to the murein [8], they constitute the most abundant protein in E. coli. It can be assumed that certain properties of the outer membrane, such as its function as a permeability barrier against certain substrates and antibiotics, are determined substantially by this lipoprotein. Knowledge of the lipoprotein's conformation would aid in the construction of a tentative model of outer membrane structure.Immunological studies with the aim of localizing the lipoprotein within the outer membrane showed that lipoprotein acts as an antigen in mutant cells with defects in cell surface structures and that in wild-type cells it is buried in the membrane (9 -111. Since the amino acid sequence of the lipoprotein is known [12,13], we could study the regions of the polypeptide chain against which antibodies were directed. From the two lipoprotein samples used for the conformational studies in this paper, one was released from the murein by degradation of murein with lysozyme and the other by cleaving the polypeptid...