دانلود رایگان مقاله لاتین اثر ماکرومولکولی بر پروتئین از سایت الزویر
عنوان فارسی مقاله:
اثر تراکم ماکرومولکولی بر پروتئین اتصالی میله ای کم چرب در سطح اسید آمینو تک
عنوان انگلیسی مقاله:
Effects of macromolecular crowding on a small lipid binding protein probed at the single-amino acid level
سال انتشار : 2016
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مقدمه انگلیسی مقاله:
1. Introduction
In the cell, proteins are subjected to the influence of a heterogeneous, crowded environment which modulates their stability, diffusion, conformational dynamics, and binding propensity [1e3]. The recognition that molecular crowding could affect protein function dates back to more than four decades ago and has determined an amazing spread of investigations on proteins in crowded solutions. After the first theoretical models, several computational and experimental studies have contributed to significantly advance our understanding of protein behaviour in native-like environments [1,4e8]. For example, in spite of the observation that the overall impact of a cellular environment on protein chemistry can be moderate [4,9], distinct forces were recognized to operate simultaneously, often producing opposite effects [10e12]. Continued efforts have recently led to the concept of structured crowding [13] and to the description of crowding-induced perturbations to solvent properties [14]. However we note that most experimental studies on globular proteins have been focused on macroscopic properties such as stability, rotational/translational diffusion, and binding affinity, while local effects of crowding have been largely disregarded. Since polypeptide functions are intimately linked to the structural and dynamic features of active sites, surface recognition patches, ligand binding pockets, or hot spot regions, it is important to assess how internal motions and local structure are perturbed in a cell-like environment. Apart from computational approaches, few techniques are available to detect the perturbations produced by a crowded environment on proteins at the single amino acid level. Among these, NMR spectroscopy plays a prominent role [15]. An arsenal of solution NMR methods allows the study of structure, dynamics, and binding of biomolecules in a broad range of time scales [16]. While * Corresponding author. suffering from unfavorable consequences of high solution viscosity in a crowded solution [17], a strength of the technique is that, after isotope-labeling, the signals of a tracer molecule can be monitored without interference from a complex background such as the cytoplasm [18,19]. 15N-enriched protein molecules can be detected through two-dimensional heteronuclear correlation (HSQC) spectra which display distinct signals from the majority of amino acid residues. Individual resonances contain information on local structure and dynamics, as well as on binding-induced perturbations. Accordingly, peak position changes were reported to reveal protein-crowder interactions [20], signal line widths were exploited to determine the viscosity of the cell interior [21], 15N- and 13Cspin relaxation rate measurements were found to be able to detect weak unspecific protein binding under crowding conditions [22,23], and NMR-detected amide proton exchange experiments enabled the evaluation of the effects of macromolecular crowding on protein stability [24]. With notable exceptions, previous work was prevalently concerned with the influence of crowding on global molecular properties such as thermodynamic stability and diffusive dynamics. However, the extent to which this phenomenon affects local structure, dynamics and stability of biopolymers as well as its impact on binding site organization, remains elusive. Herein, we report an NMR-based study aimed at a systematic evaluation of the effects of cell-mimicking chemistry on individual amino acids along the polypeptide backbone of the liver fatty acid binding protein (LFABP). LFABP belongs to a group of abundantly expressed cytosolic proteins that have been implicated in intracellular lipid trafficking, lipid signal regulation, and systemic metabolic homeostasis [25e27]. These proteins, called intracellular lipid binding proteins (iLBPs), consist of a ten-stranded antiparallel b-barrel and a short helix-turn-helix motif surrounding an internal ligand-binding cavity [28]. The hollow structure and the measurable localized internal dynamics of the ligand-free form of iLBPs in dilute solution [28e30] suggest that they may be more susceptible to crowding-induced perturbations than a compact globular protein. Upon fatty acid binding, LFABP largely retains its tertiary structure and occupies a similar volume to that occupied by the free protein (21.33 nm3 and 21.07 nm3 , respectively). However, the density of non-solvent atoms in the protein-ligand complex is enhanced compared to the unbound form, and a bound liganddependent dynamics has been reported [31,32]. Therefore, LFABP represents an intriguing model to test if a small dynamic protein is affected by macromolecular cosolutes as well as to investigate crowding-induced perturbations to the mechanism of small molecule binding.
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کلمات کلیدی:
[PDF]Effects of macromolecular crowding on a small lipid binding protein ... iranarze.ir/wp-content/uploads/2016/10/E346.pdf Jul 22, 2016 - Effects of macromolecular crowding on a small lipid binding protein probed at the single-amino acid level. Silvia Pérez Santero, Filippo Favretto ... Quadruplex Nucleic Acids - Page 110 - Google Books Result https://books.google.com/books?isbn=3642347436 Jonathan B. Chaires, David Graves - 2014 - Science ... Sugimoto N (2006) Effect of molecular crowding on DNA polymerase activity. ... telomerase by small chemical ligands under molecular crowding condition. ... Wang J, Zhang M (2010) Spectroscopic study on the binding of porphyrins to ... crowding agents improve passive biomacromolecule encapsulation in lipid vesicles. Protein Misfolding, Aggregation and Conformational Diseases: Part B: ... https://books.google.com/books?isbn=0387365346 Vladimir N. Uversky, Anthony Fink - 2007 - Medical Such reversible lipid membrane binding is suggested by the sequence of the protein, which ... Effect of Molecular Crowding The concentration of macromolecules, including proteins, nucleic acids, carbohydrates, and small solutes, within a ... [PDF]Macromolecular crowding: obvious but underappreciated www.rpgroup.caltech.edu/courses/aph161/Handouts/Ellis2001B.pdf by RJ Ellis - 2001 - Cited by 1572 - Related articles effects of crowding raises doubts about the relevance ... dispersed in small elements or pores, with dimensions comparable to the size of large ... sufficient to cause significant crowding effects .... volume obtained when macromolecules bind to one another. ...... lipid-binding sites in cytoskeletal proteins have been identified. Macromolecular Crowding Effects on Multiprotein Binding Equilibria ... www.cell.com/biophysj/pdf/S0006-3495(10)05040-X.pdf Mar 9, 2011 - of lipid-mediated contributions to such effects with major implications on pro ... simulation in better exploring the energy landscape of a small protein under ... Macromolecular Crowding Effects on Multiprotein Binding Equilibria ... Molecular Crowding Drives Active Pin1 into Nonspecific Complexes ... pubs.acs.org/doi/abs/10.1021/ja405244v by LM Luh - 2013 - Cited by 34 - Related articles Aug 22, 2013 - In addition, we demonstrate that commonly used synthetic crowding .... Effects of macromolecular crowding on a small lipid binding protein ... [PDF]Effect of Macromolecular Crowding on Protein Folding Dynamics at ... https://www.med.upenn.edu/shorterlab/Papers/.../1-s2.0-S0022283609010006-main.p... by L Guo - 2009 - Cited by 67 - Related articles Aug 13, 2009 - into the effect of crowding on the stability and folding rate of protein tertiary structures ... small folding motifs (i.e., a 34-residue α-helix, a 34-residue cross-linked helix–turn–helix ..... structural motif found in DNA-binding proteins.46. We have ..... Heterogeneous and anomalous diffusion inside lipid tubules.
