Supplementary Figure 8: Distance restraint and distribution on the 3D structure of rdHDL. | Nature Structural & Molecular Biology

Supplementary Figure 8: Distance restraint and distribution on the 3D structure of rdHDL.

From: Solution structure of discoidal high-density lipoprotein particles with a shortened apolipoprotein A-I

Supplementary Figure 8

(a) [1H,13C] HMQC of the methyl region showing the proS methyls of Leucine and Valines (note that the sequence does not contain Isoleucines). Selective labeling was achieved following the approach from the Boisbouvier group (see Materials and Methods section for more details). (b) 2D plane of a 3D CHH-NOESY-HMQC using the methyl-selective labeled sample to determine intra- and intermolecular methyl-methyl NOEs. (c) Excerpt from 2D [1H,15N]-TROSY spectra which were measured to determine intermolecular PRE effects. In order to do so, 14N-1H-MTSL-Cys-variant-MSPΔH5 (here in this case 213,192,148) and 15N-2H-wildtype-MSPΔH5 were assembled. Since only the 15N-2H-wildtype-MSPΔH5 is NMR-visible which has no MTSL tag, signal broadening must stem from the other NMR-invisible but MTSL-tagged molecule and therefore intermolecular. As it can be seen, several residues experience intermolecular PRE broadening. (d) Long range distance restraints (blue lines) collected from PRE data and NOEs are mapped onto the 3D structure of rdHDL.

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