In Phase 1, mice were reinforced for nose poking on a fixed-ratio (FR1) schedule of reinforcement on either the right or left side (assignment of reinforced hole counterbalanced across groups).
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We repeated measurements of female association times (including switching of side-assignments of the two stimulus males) as described for the 1st part of the experiments.
The user interface accommodates the six phases of the STRIP medication review (i.e. drugs disease assignment, undertreatment, overtreatment, side effects drugs assignment, clinical interactions and dosage frequency).
Experiments using 2D 1H-15N and 1H-13C HSQC, 3D HNCA, CBCA CO NH, HNCACB, HNCO, HN CA CO, HBHA CO NH, HBHANH, HCCH-TOCSY, and CCH-TOCSY were performed for the backbone and side chain assignments of Rpn13C.
3D 15N-edited NOESY-HSQC, 4D 13C,15N-edited NOESY and 4D 13C,13C-edited NOESY experiments were used for side chain assignments and to identify long range intra- and interdomain [1H,1H] NOEs [30].
For V22-SH3, NMR spectra were acquired for both backbone and side chain assignments which included 15N-edited HSQC-TOCSY, HSQC-NOESY and 13C-HCCH-TOCSY as well as triple-resonance experiments [HNCACB, CBCA CO NH, HNCO].
These tools, along with chemical shift predictions from the PACSY database, greatly accelerate protein side chain assignments.
Methyl side chain assignments for several intermediates of the hDHFR major enzymatic cycle are summarized in Table S1 of the Supporting Information.
The electron density of the peptide is well defined at this resolution, so there is no ambiguity regarding the direction of the polypeptide nor the visible side chain assignments.
Summary: ADAPT-nuclear magnetic resonance (ADAPT-NMR) offers an automated approach to the concurrent acquisition and processing of protein NMR data with the goal of complete backbone and side chain assignments.
