Translational and rotational motions of proteins in a protein crowded environment

Zorrilla, Silvia; Hink, Mark A.; Visser, Antonie J. W. G.; Lillo, M. Pilar

doi:10.1016/j.bpc.2006.09.003

Cited by 60 publications

(82 citation statements)

References 48 publications

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“…Similar concentration dependences of rotational and translational protein diffusion have been reported previously (4), but this is the first time, to our knowledge, that such a large quantitative difference has been observed in a protein system. The effect of less hindered rotations compared to the translational self-diffusion goes far beyond a pure viscosity effect resulting from the difference in local microviscosity around the protein and the bulk viscosity.…”

Section: The Impact Of Crowding: Rotational Diffusion Is Less Hinderesupporting

confidence: 77%

NMR-Detected Brownian Dynamics of α B-Crystallin over a Wide Range of Concentrations

Roos

Link

Balbach

et al. 2015

Biophysical Journal

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Knowledge about the global translational and rotational motion of proteins under crowded conditions is highly relevant for understanding the function of proteins in vivo. This holds in particular for human αB-crystallin, which is strongly crowded in vivo and inter alia responsible for preventing cataracts. Quantitative information on translational and rotational diffusion is not readily available, and we here demonstrate an approach that combines pulsed-field-gradient NMR for translational diffusion and proton T1ρ/T2 relaxation-time measurements for rotational diffusion, thus overcoming obstacles encountered in previous studies. The relaxation times measured at variable temperature provide a quantitative measure of the correlation function of protein tumbling, which cannot be approximated by a single exponential, because two components are needed for a minimal and adequate description of the data. We find that at high protein concentrations, rotational diffusion is decoupled from translational diffusion, the latter following the macroscopic viscosity change almost quantitatively, resembling the behavior of spherical colloids. Analysis of data reported in the literature shows that well-packed globular proteins follow a scaling relation between the hydrodynamic radius and the molar mass, Rh ∼ M(1/d), with a fractal dimension of d ∼ 2.5 rather than 3. Despite its oligomeric nature, Rh of αB-crystallin as derived from both NMR methods is found to be fully consistent with this relation.

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Section: The Impact Of Crowding: Rotational Diffusion Is Less Hinderesupporting

confidence: 77%

NMR-Detected Brownian Dynamics of α B-Crystallin over a Wide Range of Concentrations

Roos

Link

Balbach

et al. 2015

Biophysical Journal

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“…It has been confirmed experimentally by Lavalette and colleagues, who observed a considerably lesser decrease of rotational diffusivity of serum albumin with increasing 17.5kDa dextran concentration than for translational diffusion of this protein (Lavalette et al, 2006). Similarly, Zorrilla and colleagues reported that the rotational diffusivity of equine apo-myoglobin depends much less on increasing concentrations of RNAse A or human serum albumin than does its translational diffusivity (Zorrilla et al, 2007). It can be concluded that the microviscosity sensed by rotational diffusion at higher protein concentrations is considerably lower than that sensed by translational diffusion.…”

Section: Insights Into Cytoplasmic Microviscosity and Architecture Frmentioning

confidence: 75%

Myoglobin's old and new clothes: from molecular structure to function in living cells

Gros

Wittenberg

Jue

2010

Journal of Experimental Biology

102

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SUMMARY Myoglobin, a mobile carrier of oxygen, is without a doubt an important player central to the physiological function of heart and skeletal muscle. Recently, researchers have surmounted technical challenges to measure Mb diffusion in the living cell. Their observations have stimulated a discussion about the relative contribution made by Mb-facilitated diffusion to the total oxygen flux. The calculation of the relative contribution, however, depends upon assumptions, the cell model and cell architecture, cell bioenergetics, oxygen supply and demand. The analysis suggests that important differences can be observed whether steady-state or transient conditions are considered. This article reviews the current evidence underlying the evaluation of the biophysical parameters of myoglobin-facilitated oxygen diffusion in cells, specifically the intracellular concentration of myoglobin, the intracellular diffusion coefficient of myoglobin and the intracellular myoglobin oxygen saturation. The review considers the role of myoglobin in oxygen transport in vertebrate heart and skeletal muscle, in the diving seal during apnea as well as the role of the analogous leghemoglobin of plants. The possible role of myoglobin in intracellular fatty acid transport is addressed. Finally, the recent measurements of myoglobin diffusion inside muscle cells are discussed in terms of their implications for cytoarchitecture and microviscosity in these cells and the identification of intracellular impediments to the diffusion of proteins inside cells. The recent experimental data then help to refine our understanding of Mb function and establish a basis for future investigation.

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“…(4)). Our study also reveals that apoMb rotational diffusive motions were less slowed down than translational ones [21]. This result can be understood on the basis of the different nature of rotational and translational diffusive motions.…”

Section: Viscosity Vs Microviscositymentioning

confidence: 77%

“…It was pointed out that the extent to which translational diffusive motions are hindered, in solutions containing high concentrations of dextrans or ficolls, strongly depends on the crowder agent used, but not on the size of the diffusing species. We have also used this method to monitor translational diffusion of apoMb in protein crowded solutions (see below) [21]. Besides, translational diffusion of various tracer species, including small fluorophores, proteins and polymers, in moderately concentrated poly(vinyl alcohol) solutions, has also been investigated by FCS [24].…”

Section: Fluorescence Correlation and Cross-correlation Spectroscopymentioning

confidence: 99%

“…Thus, certain crowding conditions have been shown to enhance spectrin tetramerization [14,15], accelerate amyloid fibril formation by -synuclein [16][17][18], and stabilize lysozyme versus thermal denaturation [19], to name some examples. Besides, steric repulsion results in significant hindrance to rotational and translational diffusive motions [20][21][22][23][24]. In consequence, macromolecular crowding may influence reaction kinetics as well as crucial phenomena such as metabolism and cellular signaling [8,25].…”

Section: Introductionmentioning

confidence: 99%

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Quantitative Investigation of Biomolecular Interactions in Crowded Media by Fluorescence Spectroscopy, a Good Choice

Zorrilla

Lillo²

2009

CPPS

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Fluorescence spectroscopy methods have been proved to be powerful tools for the quantitative investigation of homologous and heterologous interactions, rotational and translational diffusion, and structural dynamics of biological molecules in crowded media. In addition to their high sensitivity, these methods present the advantage that the selective fluorescent labeling of the biomolecules under study allows distinguishing them from the background species. Moreover, the recent development of biological applications of single molecule fluorescence micro-spectroscopy methods has opened the possibility of performing quantitative determinations inside cells. In the last decades, theoretical and experimental studies have demonstrated the possible influence of the high concentration of macromolecules within living systems, on the thermodynamics and kinetics of biological reactions. Therefore, there is a growing interest in quantitatively evaluating the interactions involving biomolecules in the natural environment in which they occur. Since this is not always feasible, experiments conducted in model crowded conditions, resembling physiological media, may contribute to reduce the gap between traditional in vitro and in vivo experiments. In this review we will discuss the application of some fluorescence spectroscopy approaches, for the identification and quantification of biological macromolecules and their functional interactions in model crowded conditions.

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Translational and rotational motions of proteins in a protein crowded environment

Cited by 60 publications

References 48 publications

NMR-Detected Brownian Dynamics of α B-Crystallin over a Wide Range of Concentrations

NMR-Detected Brownian Dynamics of α B-Crystallin over a Wide Range of Concentrations

Myoglobin's old and new clothes: from molecular structure to function in living cells

Quantitative Investigation of Biomolecular Interactions in Crowded Media by Fluorescence Spectroscopy, a Good Choice

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