Tissue Factor/Factor VIIa Complex: Role of the Membrane Surface

Morrissey, James H.; Tajkhorshid, Emad; Sligar, Stephen G.; Rienstra, Chad M.

doi:10.1016/j.thromres.2012.02.019

Cited by 25 publications

(20 citation statements)

References 30 publications

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“…The coagulation factor X binding to Nanodiscs assembled with POPC:POPS mixtures of different compositions, from 0 to 90% PS lipid, showed that maximum affinity is reached at 80% PS, and the maximum rate of Factor X activation by the complex of tissue factor and Factor VIIa is observed at 70% PS 91 . This observation confirmed the idea of preferential activation of Factor X on PS-rich clusters, the formation of which may be favored by the presence of divalent cations 263 .…”

Section: Application Of Nanodiscs In Functional Studies Of Membransupporting

confidence: 85%

Nanodiscs in Membrane Biochemistry and Biophysics

2017

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Membrane proteins play a most important part in metabolism, signaling, cell motility, transport, development, and many other biochemical and biophysical processes which constitute fundamentals of life on molecular level. Detailed understanding of these processes is necessary for the progress of life sciences and biomedical applications. Nanodiscs provide a new and powerful tool for a broad spectrum of biochemical and biophysical studies of membrane proteins and are commonly acknowledged as an optimal membrane mimetic system which provides control over size, composition and specific functional modifications on nanometer scale. In this review we attempted to combine a comprehensive list of various applications of Nanodisc technology with systematic analysis of the most attractive features of this system and advantages provided by Nanodiscs for structural and mechanistic studies of membrane proteins.

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Section: Application Of Nanodiscs In Functional Studies Of Membransupporting

confidence: 85%

Nanodiscs in Membrane Biochemistry and Biophysics

2017

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“…Beyond providing a simple system for assessing Nanodisc assembly, “empty” Nanodiscs of precise lipid composition have found broad utility in probing protein-lipid interactions of fundamental importance to several biological processes, including the blood coagulation cascade. 43–45 Nanodiscs were formed using either 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) or 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), and two different MSPs (MSP1D1 or MSP1E3D1). MSP1D1 results in Nanodiscs 9.7 nm in diameter with 120 to 160 lipids per Nanodisc and a lipid:MSP ratio of 60:1 to 80:1 (there are two MSPs per Nanodisc), depending on the packing density of the lipids.…”

Section: Resultsmentioning

confidence: 99%

Microfluidic platform for efficient Nanodisc assembly, membrane protein incorporation, and purification

et al. 2017

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The characterization of integral membrane proteins presents numerous analytical challenges on account of their poor activity under non-native conditions, limited solubility in aqueous solutions, and low expression in most cell culture systems. Nanodiscs are synthetic model membrane constructs that offer many advantages for studying membrane protein function by offering a native-like phospholipid bilayer environment. The successful incorporation of membrane proteins within Nanodiscs requires experimental optimization of conditions. Standard protocols for Nanodisc formation can require large amounts of time and input material, limiting the facile screening of formation conditions. Capitalizing on the miniaturization and efficient mass transport inherent to microfluidics, we have developed a microfluidic platform for efficient Nanodisc assembly and purification, and demonstrated the ability to incorporate functional membrane proteins into the resulting Nanodiscs. In addition to working with reduced sample volumes, this platform simplifies membrane protein incorporation from a multi-stage protocol requiring several hours or days into a single platform that outputs purified Nanodiscs in less than one hour. To demonstrate the utility of this platform, we incorporated Cytochrome P450 into Nanodiscs of variable size and lipid composition, and present spectroscopic evidence for the functional active site of the membrane protein. This platform is a promising new tool for membrane protein biology and biochemistry that enables tremendous versatility for optimizing the incorporation of membrane proteins using microfluidic gradients to screen across diverse formation conditions.

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“…Findings of these studies have been reviewed recently by Morrissey et al. [58,59] and will not be discussed in detail here unless they are relevant for understanding how PS could regulate TF‐FVIIa activity at the cell surface. Although studies in artificial membranes with defined lipid content are useful for understanding the potential effects of phospholipids on TF activity, they may not accurately reflect the behavior of complex cell membrane surfaces.…”

Section: Potential Mechanisms By Which Ps Regulates Tf‐fviia Coagulanmentioning

confidence: 99%

Regulation of tissue factor coagulant activity on cell surfaces

Rao

Pendurthi

2012

Journal of Thrombosis and Haemostasis

102

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Summary Tissue factor (TF) is a transmembrane glycoprotein and an essential component of factor VIIa-TF enzymatic complex that triggers activation of the coagulation cascade. Formation of TF-FVIIa complexes on cell surfaces not only trigger the coagulation cascade but also transduce cell signaling via activation of protease-activated receptors. Tissue factor is expressed constitutively on cell surfaces of a variety of extravascular cell types, including fibroblasts and pericytes in and surrounding blood vessel walls and epithelial cells but generally absent on cells that come in contact with blood directly. However, TF expression could be induced in some blood cells, such as monocytes and endothelial cells, following an injury or pathological stimuli. Tissue factor is essential for hemostasis, but aberrant expression of TF leads to thrombosis. Therefore, a proper regulation of TF activity is critical for the maintenance of hemostatic balance and health in general. TF-FVIIa coagulant activity at the cell surface is influenced not only by TF protein expression levels but also independently by a variety of mechanisms, including alterations in membrane phospholipid composition and cholesterol content, thiol-dependent modifications of TF allosteric disulfide bond, and other post-translational modifications of TF. In this article, we critically review key literature on mechanisms by which TF coagulant activity is regulated at the cell surface in the absence of changes in TF protein levels with specific emphasis on recently published data and provide the authors’ perspective on the subject.

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Tissue Factor/Factor VIIa Complex: Role of the Membrane Surface

Cited by 25 publications

References 30 publications

Nanodiscs in Membrane Biochemistry and Biophysics

Nanodiscs in Membrane Biochemistry and Biophysics

Microfluidic platform for efficient Nanodisc assembly, membrane protein incorporation, and purification

Regulation of tissue factor coagulant activity on cell surfaces

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