In Situ Solid‐State NMR Spectroscopy of Protein in Heterogeneous Membranes: The Baseplate Antenna Complex of <i>Chlorobaculum tepidum</i>

Kulminskaya, Natalia; Pedersen, Marie Østergaard; Bjerring, Morten; Underhaug, Jarl; Miller, Megan; Frigaard, Niels-Ulrik; Nielsen, Jakob T.; Nielsen, Niels Chr.

doi:10.1002/ange.201201160

Cited by 3 publications

(7 citation statements)

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“…In accord with indications from earlier studies, the CsmA monomers form helical structures with a slight curvature20. The data presented here reveal that the helices form rods by head-to-tail translational association in a tight interface allowed by the small amino acids of in particular G15, G39 and A12 flanked by other relatively small polar amino acids such as T8, S35 and N46 (Figs 3d and 4c).…”

Section: Resultssupporting

confidence: 91%

“…From our previous solid-state NMR investigations, we could, based on secondary chemical shift data, conclude that the CsmA monomers in situ in the baseplate display a largely α-helical structure encapsulated into a fully symmetric baseplate super-structure20. Furthermore, we conclude now, based on analysis of H25 side chain chemical shifts (Supplementary Note 2) that BChl a ligands are coordinated 1:1 via H25.…”

Section: Resultsmentioning

confidence: 65%

“…In detail, 2D homonuclear 13 C- 13 C DARR, heteronuclear NCA and NCO experiments with broad-banded version as well as 3D NCACX, NCOCX, CONCA, CANCO spectra were obtained. Details for those experiments have been described previously20. The 2D homonuclear 13 C– 13 C DARR62 correlation spectrum (shown in Supplementary Fig.…”

Section: Methodsmentioning

confidence: 99%

“…tepidum , removing CsmA and thus the baseplate cannot give viable mutants18. Initially, we presented a liquid-state NMR structure of CsmA in organic solution19 and later extended the investigation with solid-state NMR resonance (ssNMR) assignments and secondary structure information20 for the CsmA peptides residing in situ in the baseplate of the carotenosome15.…”

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confidence: 99%

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In situ high-resolution structure of the baseplate antenna complex in Chlorobaculum tepidum

et al. 2016

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Photosynthetic antenna systems enable organisms harvesting light and transfer the energy to the photosynthetic reaction centre, where the conversion to chemical energy takes place. One of the most complex antenna systems, the chlorosome, found in the photosynthetic green sulfur bacterium Chlorobaculum (Cba.) tepidum contains a baseplate, which is a scaffolding super-structure, formed by the protein CsmA and bacteriochlorophyll a. Here we present the first high-resolution structure of the CsmA baseplate using intact fully functional, light-harvesting organelles from Cba. tepidum, following a hybrid approach combining five complementary methods: solid-state NMR spectroscopy, cryo-electron microscopy, isotropic and anisotropic circular dichroism and linear dichroism. The structure calculation was facilitated through development of new software, GASyCS for efficient geometry optimization of highly symmetric oligomeric structures. We show that the baseplate is composed of rods of repeated dimers of the strongly amphipathic CsmA with pigments sandwiched within the dimer at the hydrophobic side of the helix.

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Section: Resultssupporting

confidence: 91%

Section: Resultsmentioning

confidence: 65%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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In situ high-resolution structure of the baseplate antenna complex in Chlorobaculum tepidum

et al. 2016

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“…In‐cell solid‐state MAS NMR may therefore be a useful tool to study the structure and dynamics of soluble large protein complexes in cells [23]. Another advance is the recently shown feasibility of using solid‐state MAS NMR for detecting membrane proteins in native cell membranes [22,52–54] and living E. coli cells [18]. Background signals can be suppressed by advanced isotope labeling strategies and more elaborate multi‐dimensional experiments.…”

Section: Perspectivesmentioning

confidence: 99%

Protein dynamics in living cells studied by in‐cell NMR spectroscopy

Liu

2013

FEBS Letters

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a b s t r a c tMost proteins function in cells where protein concentrations can reach 400 g/l. However, most quantitative studies of protein properties are performed in idealized, dilute conditions. Recently developed in-cell NMR techniques can provide protein structure and other biophysical properties inside living cells at atomic resolution. Here we review how protein dynamics, including global and internal motions have been characterized by in-cell NMR, and then discuss the remaining challenges and future directions.

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Membrane proteins in their native habitat as seen by solid‐state NMR spectroscopy

Brown

Ladizhansky

2015

Protein Science

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Membrane proteins play many critical roles in cells, mediating flow of material and information across cell membranes. They have evolved to perform these functions in the environment of a cell membrane, whose physicochemical properties are often different from those of common cell membrane mimetics used for structure determination. As a result, membrane proteins are difficult to study by traditional methods of structural biology, and they are significantly underrepresented in the protein structure databank. Solid-state Nuclear Magnetic Resonance (SSNMR) has long been considered as an attractive alternative because it allows for studies of membrane proteins in both native-like membranes composed of synthetic lipids and in cell membranes. Over the past decade, SSNMR has been rapidly developing into a major structural method, and a growing number of membrane protein structures obtained by this technique highlights its potential. Here we discuss membrane protein sample requirements, review recent progress in SSNMR methodologies, and describe recent advances in characterizing membrane proteins in the environment of a cellular membrane.

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In Situ Solid‐State NMR Spectroscopy of Protein in Heterogeneous Membranes: The Baseplate Antenna Complex of Chlorobaculum tepidum

Cited by 3 publications

References 30 publications

In situ high-resolution structure of the baseplate antenna complex in Chlorobaculum tepidum

In situ high-resolution structure of the baseplate antenna complex in Chlorobaculum tepidum

Protein dynamics in living cells studied by in‐cell NMR spectroscopy

Membrane proteins in their native habitat as seen by solid‐state NMR spectroscopy

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