Medium- and Long-Distance 1H–13C Heteronuclear Correlation NMR in Solids

Yao, Xiaolan; Schmidt‐Rohr, Klaus; Hong, Mei

doi:10.1006/jmre.2001.2285

Cited by 50 publications

(56 citation statements)

References 28 publications

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“…The resulting 2D ( 1 H-13 C)-REDOR-HETCOR spectrum contains a finite number of well-resolved intermolecular cross-peaks (Fig. 4B) REDOR filters have also been used for studies of water-protein interfaces, as implemented in a medium-and long-distance MELODI-HETCOR experiment (37,38).…”

Section: Resultsmentioning

confidence: 99%

Atomic-resolution structure of the CAP-Gly domain of dynactin on polymeric microtubules determined by magic angle spinning NMR spectroscopy

Yan

Guo

Hou

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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Microtubules and their associated proteins perform a broad array of essential physiological functions, including mitosis, polarization and differentiation, cell migration, and vesicle and organelle transport. As such, they have been extensively studied at multiple levels of resolution (e.g., from structural biology to cell biology). Despite these efforts, there remain significant gaps in our knowledge concerning how microtubule-binding proteins bind to microtubules, how dynamics connect different conformational states, and how these interactions and dynamics affect cellular processes. Structures of microtubule-associated proteins assembled on polymeric microtubules are not known at atomic resolution. Here, we report a structure of the cytoskeleton-associated protein glycine-rich (CAP-Gly) domain of dynactin motor on polymeric microtubules, solved by magic angle spinning NMR spectroscopy. We present the intermolecular interface of CAP-Gly with microtubules, derived by recording direct dipolar contacts between CAP-Gly and tubulin using double rotational echo double resonance (dREDOR)-filtered experiments. Our results indicate that the structure adopted by CAP-Gly varies, particularly around its loop regions, permitting its interaction with multiple binding partners and with the microtubules. To our knowledge, this study reports the first atomic-resolution structure of a microtubule-associated protein on polymeric microtubules. Our approach lays the foundation for atomic-resolution structural analysis of other microtubule-associated motors.magic angle spinning NMR | microtubules | dynactin's CAP-Gly domain | structure determination | intermolecular interface determination

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Section: Resultsmentioning

confidence: 99%

Atomic-resolution structure of the CAP-Gly domain of dynactin on polymeric microtubules determined by magic angle spinning NMR spectroscopy

Yan

Guo

Hou

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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“…The 2D NMR experiment combines MELODI filter (24,25), LG-CP block with wPMLG3 þ decoupling (26)(27)(28) to yield medium-and long-range 1 H-13 C correlation spectra with 1 H-enhanced sensitivity. The wPMLG3 þ decoupling efficiency correlates to several experimental parameters, like rf field power (ν 1 ), unit pulse length (Δt), acquisition windows (τ w ), rotation frequency (ν r ), and 1 H rf offset frequency (Δν).…”

Section: Methodsmentioning

confidence: 99%

“…S1 and S2). The dipolar filter period of 38 μs was set to suppress the 1 H-1 H homonuclear dipolar interaction effectively (24,25). The 1 H power was ramped 80-100% during cross-polarization, and 1 H rf field strength for TPPM (two-pulse phase modulation) decoupling was about 80 kHz.…”

Section: Methodsmentioning

confidence: 99%

“…We represent 1 H sensitivity-enhanced heteronuclear NMR correlation experiment (MELODI-HETCOR) for selectively recognizing 1 H contacts of the uniformly 13 C∕ 15 N-labeled phycocyanobilin (PCB) chromophore in both Pr and Pfr states at 100% occupancy. The MELODI filter effectively silences magnetization of protons directly bonded to the chromophore, while emphasizing its medium-and long-range 1 H contacts (24,25). The sensitivity enhancement of 1 H indirect dimension was achieved through the use of windowed phase-modulated Lee-Goldburg (wPMLG) homonuclear decoupling scheme (26)(27)(28) over the 1 H evolution period (t 1 ), in our case, wPMLG3 þ .…”

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confidence: 99%

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Two ground state isoforms and a chromophore D -ring photoflip triggering extensive intramolecular changes in a canonical phytochrome

Song

Psakis

Lang

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

160

299

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Phytochrome photoreceptors mediate light responses in plants and in many microorganisms. Here we report studies using 1 H-13 C magic-angle spinning NMR spectroscopy of the sensor module of cyanobacterial phytochrome Cph1. Two isoforms of the red-light absorbing Pr ground state are identified. Conclusive evidence that photoisomerization occurs at the C15-methine bridge leading to a β-facial disposition of the ring D is presented. In the far-red-light absorbing Pfr state, strong hydrogen-bonding interactions of the D-ring carbonyl group to Tyr-263 and of N24 to Asp-207 hold the chromophore in a tensed conformation. Signaling is triggered when Asp-207 is released from its salt bridge to Arg-472, probably inducing conformational changes in the tongue region. A second signal route is initiated by partner swapping of the B-ring propionate between Arg-254 and Arg-222.chromophore-protein interaction | signal transduction | solid-state NMR | photomorphogenesis P hytochrome was first demonstrated in plants as a red-lightdependent photoreceptor regulating numerous photomorphogenic processes (1, 2). Phytochromes are, however, also now known in photosynthetic prokaryotes including cyanobacteria (3, 4), nonphotosynthetic bacteria (5), and fungi (6). Generally, the phytochrome apoprotein binds an open-chain tetrapyrrole as a chromophore (7,8) to form the red-light absorbing Pr ground state (λ max ≈ 658 nm in case of cyanobacterial phytchrome Cph1 from Synechocystis 6803). Red light absorption photoactivates the molecule to form the photoactivated far-red-light absorbing Pfr state (λ max ≈ 702 nm for Cph1) via a series of intermediates (8-10). Photoactivation is thought to be initiated by a double bond isomerization of the chromophore (10, 11). Early NMR spectroscopic studies on proteolytic phytochrome fragments (12, 13) indicated that this isomerization occurs at the C15═C16 double bond (for numbering, see Fig. 1A), a geometrical change in line with vibrational spectroscopic investigations (14-16) and results from recent 13 C solid-state NMR (17, 18) in which the most significant changes during the light-triggered conversions are confined to rings C and D. Exact geometries of the chromophore in the Pr state have been resolved as periplanar ZZZssa configurations in bacteriophytochromes from Deinococcus radiodurans (19) and Rhodopseudomonas palustris (20) as well as in the more plant-phytochrome-like Cph1 from the cyanobacterium Synechocystis 6803 (21). On the other hand, the crystal structure of the unusual bacteriophytochrome PaBphP Pseudomonas aeruginosa (22) whose ground state is Pfr shows a ZZEssa conformation, consistent with the expected primary photochemistry at the C15═C16 double bond (Fig. 1 B vs. C). Very recently, however, Ulijasz et al. presented structural simulations based on liquid NMR data of a 20-kDa GAF (cGMP phosphodiesterase/ adenylyl cyclase/FhlA) domain fragment of "SyB-Cph1" phytochrome from the thermotolerant cyanobacterium Synechococcus OSB′ (23). Surprisingly, they concluded that photoisomerization occ...

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“…Although the information can be obtained by performing the above-mentioned experiments in doubly labeled systems, the real challenge lies in obtaining long and medium range distance information in systems at natural abundance. 46 The distance obtained from such experiments based on dipolar coupling would be more accurate than the corresponding NOE-based distances in solution. Efforts toward obtaining secondary structural information on the above-mentioned peptides at natural abundance using the SSNMR techniques are under way.…”

Section: Discussionmentioning

confidence: 99%

Natural abundant solid state NMR studies in designed tripeptides for differentiation of multiple conformers

2009

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Solid state NMR (SSNMR) experiments on heteronuclei in natural abundance are described for three synthetically designed tripeptides Piv-(L)Pro-(L)Pro-(L)Phe-OMe (1), Piv-(D)Pro-(L)Pro-(L)Phe-OMe (2), and Piv-(D)Pro-(L)Pro-(L)Phe-NHMe (3). These peptides exist in different conformation as shown by solution state NMR and single crystal X-ray analysis (Chatterjee et al., Chem Eur J 2008, 14, 6192). In this study, SSNMR has been used to probe the conformations of these peptides in their powder form. The (13)C spectrum of peptide (1) showed doubling of resonances corresponding to cis/cis form, unlike in solution where the similar doubling is attributed to cis/trans form. This has been confirmed by the chemical shift differences of C(beta) and C(gamma) carbon of Proline in peptide (1) both in solution and SSNMR. Peptide (2) and (3) provided single set of resonances which represented all trans form across the di-Proline segment. The results are in agreement with the X-ray analysis. Solid state (15)N resonances, especially from Proline residues provided additional information, which is normally not observable in solution state NMR. (1)H chemical shifts are also obtained from a two-dimensional heteronuclear correlation experiment between (1)H--(13)C. The results confirm the utility of NMR as a useful tool for identifying different conformers in peptides in the solid state.

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Medium- and Long-Distance 1H–13C Heteronuclear Correlation NMR in Solids

Cited by 50 publications

References 28 publications

Atomic-resolution structure of the CAP-Gly domain of dynactin on polymeric microtubules determined by magic angle spinning NMR spectroscopy

Atomic-resolution structure of the CAP-Gly domain of dynactin on polymeric microtubules determined by magic angle spinning NMR spectroscopy

Two ground state isoforms and a chromophore D -ring photoflip triggering extensive intramolecular changes in a canonical phytochrome

Natural abundant solid state NMR studies in designed tripeptides for differentiation of multiple conformers

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