Nuclear Magnetic Resonance Structural Studies of Membrane Proteins in Micelles and Bilayers

Gong, Xiaomin; Franzin, Carla M.; Thai, Khang; Yu, Jinghua; Marassi, Francesca M.

doi:10.1007/978-1-59745-519-0_35

Cited by 17 publications

(11 citation statements)

References 58 publications

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“…Solution and solid state NMR is starting to make a significant dent, however, and it appears that GPCR-sized proteins may become accessible to these techniques. [67][68][69] For both crystallography and NMR, the key elements that can pave the way to a structure are high-level expression, protein stability, and conformational homogeneity. New methods that can attack these fundamental issues more reliably, more generally and with greater ease of implementation will continue to build the membrane protein structure revolution.…”

Section: Resultsmentioning

confidence: 99%

G‐protein‐coupled receptor structures were not built in a day

Blois

Bowie

2009

Protein Science

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Among the most exciting recent developments in structural biology is the structure determination of G-protein-coupled receptors (GPCRs), which comprise the largest class of membrane proteins in mammalian cells and have enormous importance for disease and drug development. The GPCR structures are perhaps the most visible examples of a nascent revolution in membrane protein structure determination. Like other major milestones in science, however, such as the sequencing of the human genome, these achievements were built on a hidden foundation of technological developments. Here, we describe some of the methods that are fueling the membrane protein structure revolution and have enabled the determination of the current GPCR structures, along with new techniques that may lead to future structures.

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Section: Resultsmentioning

confidence: 99%

G‐protein‐coupled receptor structures were not built in a day

Blois

Bowie

2009

Protein Science

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“…Consequently, the community of researchers has been motivated to develop the computational and mathematical methods required to fully exploit this new resource. Applications of the RDC data include the study of challenging biomolecules such as membrane proteins [1–4], carbohydrates [5–7], and DNA/RNA [8–11] to name a few. Recent advances include structure determination from multiple alignment media [12–15], reconstruction of the relative order tensors from unassigned RDC data [15,16], and identification of homologous structures [17,18] from unassigned RDC data.…”

Section: Introductionmentioning

confidence: 99%

Efficient and accurate estimation of relative order tensors from λ-maps

Mukhopadhyay¹,

Miao²,

Shealy³

et al. 2009

Journal of Magnetic Resonance

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The rapid increase in the availability of RDC data from multiple alignment media in recent years has necessitated the development of more sophisticated analyses that extract the RDC data’s full information content. This article presents an analysis of the distribution of RDCs from two media (2D-RDC data), using the information obtained from a λ-map. This article also introduces an efficient algorithm, which leverages these findings to extract the order tensors for each alignment medium using unassigned RDC data in the absence of any structural information. The results of applying this 2D-RDC analysis method to synthetic and experimental data are reported in this article. The relative order tensor estimates obtained from the 2D-RDC analysis are compared to order tensors obtained from the program REDCAT after using assignment and structural information. The final comparisons indicate that the relative order tensors estimated from the unassigned 2D-RDC method very closely match the results from methods that require assignment and structural information. The presented method is successful even in cases with small datasets. The results of analyzing experimental RDC data for the protein 1P7E are presented to demonstrate the potential of the presented work in accurately estimating the principal order parameters from RDC data that incompletely sample the RDC space. In addition to the new algorithm, a discussion of the uniqueness of the solutions is presented; no more than two clusters of distinct solutions have been shown to satisfy each λ-map.

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“…For example, in the Mycobacterium tuberculosis genome more than 50% of the 1160 putative membrane proteins have just one or two transmembrane helices; 15% of the putative membrane proteins have a molecular weight less than 20 kDa, and 36% have a molecular weight less than 30 kDa. 1 Both solution and solid-state NMR have made some progress with this important and sizeable class of proteins, 2 and just recently, the small four-helix bundle transmembrane domain of M2 proton channel from influenza A virus has been structurally characterized by X-ray crystallography, 3 by solution NMR 4 and previously by solid-state NMR. 5 Here, we describe the characterization by solution NMR of the M. tuberculosis membrane protein Rv1761c, a protein with a single transmembrane helix and a 10 kDa C-terminal domain.…”

Section: Introductionmentioning

confidence: 99%

Backbone structure of a small helical integral membrane protein: A unique structural characterization

et al. 2008

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The structural characterization of small integral membrane proteins pose a significant challenge for structural biology because of the multitude of molecular interactions between the protein and its heterogeneous environment. Here, the three-dimensional backbone structure of Rv1761c from Mycobacterium tuberculosis has been characterized using solution NMR spectroscopy and dodecylphosphocholine (DPC) micelles as a membrane mimetic environment. This 127 residue single transmembrane helix protein has a significant (10 kDa) C-terminal extramembranous domain. Five hundred and ninety distance, backbone dihedral, and orientational restraints were employed resulting in a 1.16 Å rmsd backbone structure with a transmembrane domain defined at 0.40 Å . The structure determination approach utilized residual dipolar coupling orientation data from partially aligned samples, long-range paramagnetic relaxation enhancement derived distances, and dihedral restraints from chemical shift indices to determine the global fold. This structural model of Rv1761c displays some influences by the membrane mimetic illustrating that the structure of these membrane proteins is dictated by a combination of the amino acid sequence and the protein's environment. These results demonstrate both the efficacy of the structural approach and the necessity to consider the biophysical properties of membrane mimetics when interpreting structural data of integral membrane proteins and, in particular, small integral membrane proteins.

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Nuclear Magnetic Resonance Structural Studies of Membrane Proteins in Micelles and Bilayers

Cited by 17 publications

References 58 publications

G‐protein‐coupled receptor structures were not built in a day

G‐protein‐coupled receptor structures were not built in a day

Efficient and accurate estimation of relative order tensors from λ-maps

Backbone structure of a small helical integral membrane protein: A unique structural characterization

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