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1.
Gilbert Richarme Mouadh Mihoub Julien Dairou Linh Chi Bui Thibaut Leger Aazdine Lamouri 《The Journal of biological chemistry》2015,290(3):1885-1897
Glycation is an inevitable nonenzymatic covalent reaction between proteins and endogenous reducing sugars or dicarbonyls (methylglyoxal, glyoxal) that results in protein inactivation. DJ-1 was reported to be a multifunctional oxidative stress response protein with poorly defined function. Here, we show that human DJ-1 is a protein deglycase that repairs methylglyoxal- and glyoxal-glycated amino acids and proteins by acting on early glycation intermediates and releases repaired proteins and lactate or glycolate, respectively. DJ-1 deglycates cysteines, arginines, and lysines (the three major glycated amino acids) of serum albumin, glyceraldehyde-3-phosphate dehydrogenase, aldolase, and aspartate aminotransferase and thus reactivates these proteins. DJ-1 prevented protein glycation in an Escherichia coli mutant deficient in the DJ-1 homolog YajL and restored cell viability in glucose-containing media. These results suggest that DJ-1-associated Parkinsonism results from excessive protein glycation and establishes DJ-1 as a major anti-glycation and anti-aging protein. 相似文献
2.
Saša N. Malkov Miodrag V. Živković Miloš V. Beljanski Srđan Đ. Stojanović Snežana D. Zarić 《The protein journal》2009,28(2):74-86
Using the data from Protein Data Bank the correlations of primary and secondary structures of proteins were analyzed. The
correlation values of the amino acids and the eight secondary structure types were calculated, where the position of the amino
acid and the position in sequence with the particular secondary structure differ at most 25. The diagrams describing these
results indicate that correlations are significant at distances between −9 and 10. The results show that the substituents
on Cβ or Cγ atoms of amino acid play major role in their preference for particular secondary structure at the same position
in the sequence, while the polarity of amino acid has significant influence on α-helices and strands at some distance in the
sequence. The diagrams corresponding to polar amino acids are noticeably asymmetric. The diagrams point out the exchangeability
of residues in the proteins; the amino acids with similar diagrams have similar local folding requirements.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献
3.
Proteins that assimilate particular elements were found to avoid using amino acids containing the element, which indicates
that the metabolic constraints of amino acids may influence the evolution of proteins. We suspected that low contents of carbon,
nitrogen, and sulfur may also be selected for economy in highly abundant proteins that consume large amounts of the resources
of cells. By analyzing recently available proteomic data in Escherichia coli, Saccharomyces cerevisiae, and Schizosaccharomyces pombe, we found that at least the carbon and nitrogen contents in amino acid side chains are negatively correlated with protein
abundance. An amino acid with a high number of carbon atoms in its side chain generally requires relatively more energy for
its synthesis. Thus, it may be selected against in highly abundant proteins either because of economy in building blocks or
because of economy in energy. Previous studies showed that highly abundant proteins preferentially use cheap (in terms of
energy) amino acids. We found that the carbon content is still negatively correlated with protein abundance after controlling
for the energetic cost of the amino acids. However, the negative correlation between protein abundance and energetic cost
disappeared after controlling for carbon content. Building blocks seem to be more restricted than energy. It seems that the
amino acid sequences of highly abundant proteins have to compromise between optimization for their biological functions and
reducing the consumption of limiting resources. By contrast, the amino acid sequences of weakly expressed proteins are more
likely to be optimized for their biological functions.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献
4.
Rodgers KJ Shiozawa N 《The international journal of biochemistry & cell biology》2008,40(8):1452-1466
Despite astounding diversity in their structure and function, proteins are constructed from 22 protein or ‘canonical’ amino acids. Hundreds of amino acid analogues exist; many occur naturally in plants, some are synthetically produced or can be produced in vivo by oxidation of amino acid side-chains. Certain structural analogues of the protein amino acids can escape detection by the cellular machinery for protein synthesis and become misincorporated into the growing polypeptide chain of proteins to generate non-native proteins. In this review we seek to provide a comprehensive overview of the current knowledge on the biosynthetic incorporation of amino acid analogues into proteins by mammalian cells. We highlight factors influencing their incorporation and how the non-native proteins generated can alter cell function. We examine the ability of amino acid analogues, representing those commonly found in damaged proteins in pathological tissues, to be misincorporated into proteins by cells in vitro, providing us with a useful tool in the laboratory to generate modified proteins representing those present in a wide-range of pathologies. We also discuss the evidence for amino acid analogue incorporation in vivo and its association with autoimmune symptoms. We confine the review to studies in which the synthetic machinery of cell has not been modified to accept non-protein amino acids. 相似文献
5.
Summary Adenovirus E1A and c-myc genes are known to be capable of transforming primary rat cells when they occur in combination with either polyoma middle-T or T24 Harvey-ras 1 genes. There was a low level of amino acid sequence homology between the nuclear adenovirus-12 (Ad12) E1A protein product (289 amino acids) and the c-myc protein based on optimal alignment and percentage identity. In contrast to others [Ralston R, Bishop JM (1983) Nature 306:803–806], we concluded that this low level of amino acid sequence homology was not significant, since rabies glycoprotein (RGP), which has no transforming function and localizes to the cell surface, had a similar low level of amino acid sequence homology to the c-myc protein. Furthermore, dot-matrix analysis, when used to test the overall level of amino acid sequence homology, showed no significant homology between c-myc and Ad12 E1A, E1B, or RGP. Thus, low levels of amino acid sequence homology between two proteins may not be sufficient to predict structural and functional similarities between them reliably, even if the two proteins appear to share a common function. 相似文献
6.
Abstract Folding of naturally occurring proteins has eluded a universal molecular level explanation till date. Rather, there is an abundance of diverse views on dominant factors governing protein folding. Through rigorous analyses of several thousand crystal structures, we observe that backbones of folded proteins display some remarkable invariant features. Folded proteins are characterized by spatially well-defined, distance dependent, and universal, neighborhoods of amino acids which defy any of the conventionally prevalent views. These findings present a compelling case for a newer view of protein folding which takes into account solvent mediated and amino acid shape and size assisted optimization of the tertiary structure of the polypeptide chain to make a functional protein. 相似文献
7.
Pons JI Rodríguez S Madrid C Juárez A Nieto JM 《Protein expression and purification》2004,35(2):293-297
Gene cloning in appropriate vectors followed by protein overexpression in Escherichia coli is the common means for protein purification. This approach has many advantages but also some drawbacks; one of these is that many proteins fail to achieve a soluble conformation when overexpressed in E. coli. Hha protein belongs to a family of nucleoid-associated proteins functionally related to the H-NS family of proteins. Hha-like proteins and H-NS-like proteins are able to semidirectly bind to each other. We show in this work that overexpressed Hha or HisHha protein (a functional derivative of Hha containing a 6x His tag at the amino end) from a T7-polymerase promoter in BL21 DE3 E. coli strains results in the vast majority of the protein accumulated in insoluble aggregates (inclusion bodies). We also show that tandem overexpression of HisHha and H-NS increases the solubility of HisHha and prevents the formation of inclusion bodies. Single amino acid substitutions in the HisHha protein, which impair interaction with H-NS, render insoluble protein even when tandem-expressed with H-NS, tandem expression of an insoluble protein and an interacting partner is an experimental strategy which could be useful to increase the solubility of other overexpressed proteins in E. coli. 相似文献
8.
Reactive oxygen species are formed in the body by several natural processes and by induced oxidative stress. The reactive oxygen species may react with the various biomolecules of the body, including proteins. In order to assess the impact of oxidative damage to proteins, we have tried to identify oxidized amino acids in blood proteins which might serve as biomarkers of oxidative damage. When oxidative damage is induced into bovine serum albumin by metal-catalysed oxidation systems, the aldehyde groups formed can be derivatized by fluoresceinamine (FINH2). Following acid hydrolysis of FINH2-derivatized protein, two major oxidation products, γ-glutamyl semialdehyde (GGS) and 2-amino-adipic semialdehyde (AAS), were found and identified by HPLC and MS. Isolation and identification of oxidized amino acids from homopolymers (poly-Arg,-Pro,-Lys,-Trp or -Leu) confirmed that GGS can originate from Arg or Pro, while AAS is an oxidation product of Lys. When oxidative stress was induced in rats by treatments with t-butyl hydroperoxide or acrolein, rat plasma protein levels of GGS and AAS were found to be significantly higher compared with control rats. The AAS-content in serum albumin or in total plasma proteins collected from eight different mammalian species was found to be inversely proportional to their maximum lifespan potential. The content of AAS in plasma proteins of untreated adult rats showed a positive correlation with the age of the rat. In young rats a negative correlation with age was found for both GGS and AAS. We conclude that GGS or AAS may be useful novel biomarkers of oxidative damage to proteins in vivo. 相似文献
9.
Due to advances in molecular biology the DNA sequences of structural genes coding for proteins are often known before a protein is characterized or even isolated. The function of a protein whose amino acid sequence has been deduced from a DNA sequence may not even be known. This has created greater interest in the development of methods to predict the tertiary structures of proteins. The a priori prediction of a protein's structure from its amino acid sequence is not yet possible. However, since proteins with similar amino acid sequences are observed to have similar three-dimensional structures, it is possible to use an analogy with a protein of known structure to draw some conclusions about the structure and properties of an uncharacterized protein. The process of predicting the tertiary structure of a protein relies very much upon computer modeling and analysis of the structure. The prediction of the structure of the bacteriophage 434 cro repressor is used as an example illustrating current procedures. 相似文献
10.
Summary The frequencies of substitutions resulting in protein instability were calculated by a method estimating changes in stability produced by amino acid substitutions. The method takes into account the accessibility of an amino acid position to a solvent and changes in the specificity of amino acid interactions. When tested on human mutant hemoglobins, the method yielded predictions with a preciseness of 80%. The consideration of the evolutionary homologous proteins in the analysis allowed us to estimate the evolutionary constraints imposed on stability of their spatial structure. With these limitations, approximately 50% of amino acid substitutions in the entire mutational spectra of the - and -subunits of human hemoglobin were found to damage the spatial structure of the globular proteins. 相似文献
11.
Tail-anchored proteins are a group of membrane proteins oriented with their amino terminus in the cytoplasm and their carboxy
terminus embedded in intracellular membranes. This group includes the apoptosis-mediating proteins of the Bcl-2 family as
well as the vesicle targeting proteins of the SNARE group, among others. A stretch of hydrophobic amino acids at the extreme
carboxy terminus of these proteins serves both as a membrane anchor and as a targeting signal. Tail-anchored proteins are
differentially targeted to either the endoplasmic reticulum or the mitochondrial outer membrane and the mechanism which accomplishes
this selective targeting is poorly understood. Here we define important characteristics of the signal/anchor region which
directs proteins to the mitochondrial outer membrane. We have created an artificial sequence consisting of a stretch of 16
leucines bounded by positively charged amino acids. Using this template we demonstrate that moderate hydrophobicity distinguishes
the mitochondrial tail-anchor sequence from that of the endoplasmic reticulum tail-anchor sequence. A change as small as introduction
of a single polar residue into a sequence that otherwise targets to the endoplasmic reticulum can substantially switch targeting
to the mitochondrial outer membrane. Further we show that a mitochondrially targeted tail-anchor has a higher propensity for
the formation of alpha-helical structure than a sequence directing tail-anchored proteins to the endoplasmic reticulum. 相似文献
12.
The amino acid composition of human alcohol dehydrogenase (ADH) was compared with alcohol dehydrogenases from different organisms
and with other proteins. Similar amino acid sequences in human ADH (template protein) and in other proteins were determined
by means of an original computer program. Analysis of amino acid motifs reveals that the ADHs from evolutionary more close
organisms have more common amino acid sequences. The quantity measure of amino acid similarity was the number of similar motifs
in analyzed protein per protein length. This value was measured for ADHs and for different proteins. For ADHs, this quotient
was higher than for proteins with different functions; for vertebrates it correlated with evolutionary closeness. The similar
operation of motif comparison was made with the help of program complex “MEME”. The analysis of ADHs revealed 4 motifs common
to 6 of 10 tested organisms and no such motifs for proteins of different function. The conclusion is that general amino composition
is more important for protein function than amino acid order and for enzymes of similar function it better correlates with
evolutionary distance between organisms. 相似文献
13.
We report here the identification of a novel domain - DIM (N-terminal domain in bacterial membrane proteins and other proteins) present exclusively in bacterial species including mycobacteria, revealed by PSI-BLAST iterative searches. DIM comprises about 53 amino acids in length with conserved Leu, Ile and Gly residues. Secondary structure prediction indicated that this domain contains two alpha-helices. DIM occurs at the N-terminus of proteins, and was found particularly but not exclusively in proteins with a transmembrane domain, and also in proteins with a FHA domain or RPT repeats. DIM-containing proteins have been reported to be involved in pathogenicity, signal transduction or small solute transport. 相似文献
14.
Summary Chou-Fasman parameters, measuring preferences of each amino acid for different conformational regions in proteins, were used to obtain an amino acid difference index of conformational parameter distance (CPD) values. CPD values were found to be significantly lower for amino acid exchanges representing in the genetic code transitions of purines, GA than for exchanges representing either transitions of pyrimidines, CU, or transversions of purines and pyrimidines. Inasmuch as the distribution of CPD values in these non GA exchanges resembles that obtained for amino acid pairs with double or triple base differences in their underlying codons, we conclude that the genetic code was not particularly designed to minimize effects of mutation on protein conformation. That natural selection minimizes these changes, however, was shown by tabulating results obtained by the maximum parsimony method for eight protein genealogies with a total occurrence of 4574 base substitutions. At the beginning position of the codons GA transitions were in very great excess over other base substitutions, and, conversely, CU transitions were deficient. At the middle position of the codons only fast evolving proteins showed an excess of GA transitions, as though selection mainly preserved conformation in these proteins while weeding out mutations affecting chemical properties of functional sites in slow evolving proteins. In both fast and slow evolving proteins the net direction of transitions and transversions was found to be from G beginning codons to non-G beginning codons resulting in more commonly occurring amino acids, especially alanine with its generalized conformational properties, being replaced at suitable sites by amino acids with more specialized conformational and chemical properties. Historical circumstances pertaining to the origin of the genetic code and the nature of primordial proteins could account for such directional changes leading to increases in the functional density of proteins.In order to further explore the course of protein evolution, a modified parsimony algorithm was developed for constructing protein genealogies on the basis of minimum CPD length. The algorithm's ability to judge with finer discrimination that in protein evolution certain pathways of amino acid substitution should occur more readily than others was considered a potential advantage over strict maximum parsimony. In developing this CPD algorithm, the path of minimum CPD length through intermediate amino acids allowed by the genetic code for each pair of amino acids was determined. It was found that amino acid exchanges representing two base changes have a considerably lower average CPD value per base substitution than the amino acid exchanges representing single base changes. Amino acid exchanges representing three base changes have yet a further marked reduction in CPD per base change. This shows how extreme constraining effects of stabilizing selection can be circumvented, for by way of intermediate amino acids almost any amino acid can ultimately be substituted for another without damage to an evolving protein's conformation during the process. 相似文献
15.
Reactive oxygen species are formed in the body by several natural processes and by induced oxidative stress. The reactive oxygen species may react with the various biomolecules of the body, including proteins. In order to assess the impact of oxidative damage to proteins, we have tried to identify oxidized amino acids in blood proteins which might serve as biomarkers of oxidative damage. When oxidative damage is induced into bovine serum albumin by metal-catalysed oxidation systems, the aldehyde groups formed can be derivatized by fluoresceinamine (FINH2). Following acid hydrolysis of FINH2-derivatized protein, two major oxidation products, γ-glutamyl semialdehyde (GGS) and 2-amino-adipic semialdehyde (AAS), were found and identified by HPLC and MS. Isolation and identification of oxidized amino acids from homopolymers (poly-Arg,-Pro,-Lys,-Trp or -Leu) confirmed that GGS can originate from Arg or Pro, while AAS is an oxidation product of Lys. When oxidative stress was induced in rats by treatments with t-butyl hydroperoxide or acrolein, rat plasma protein levels of GGS and AAS were found to be significantly higher compared with control rats. The AAS-content in serum albumin or in total plasma proteins collected from eight different mammalian species was found to be inversely proportional to their maximum lifespan potential. The content of AAS in plasma proteins of untreated adult rats showed a positive correlation with the age of the rat. In young rats a negative correlation with age was found for both GGS and AAS. We conclude that GGS or AAS may be useful novel biomarkers of oxidative damage to proteins in vivo. 相似文献
16.
Remote homology detection refers to the detection of structure homology in evolutionarily related proteins with low sequence similarity. Supervised learning algorithms such as support vector machine (SVM) are currently the most accurate methods. In most of these SVM-based methods, efforts have been dedicated to developing new kernels to better use the pairwise alignment scores or sequence profiles. Moreover, amino acids’ physicochemical properties are not generally used in the feature representation of protein sequences. In this article, we present a remote homology detection method that incorporates two novel features: (1) a protein's primary sequence is represented using amino acid's physicochemical properties and (2) the similarity between two proteins is measured using recurrence quantification analysis (RQA). An optimization scheme was developed to select different amino acid indices (up to 10 for a protein family) that are best to characterize the given protein family. The selected amino acid indices may enable us to draw better biological explanation of the protein family classification problem than using other alignment-based methods. An SVM-based classifier will then work on the space described by the RQA metrics. The classification scheme is named as SVM-RQA. Experiments at the superfamily level of the SCOP1.53 dataset show that, without using alignment or sequence profile information, the features generated from amino acid indices are able to produce results that are comparable to those obtained by the published state-of-the-art SVM kernels. In the future, better prediction accuracies can be expected by combining the alignment-based features with our amino acids property-based features. Supplementary information including the raw dataset, the best-performing amino acid indices for each protein family and the computed RQA metrics for all protein sequences can be downloaded from http://ym151113.ym.edu.tw/svm-rqa. 相似文献
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19.
Acute Effect of Ammonia on Branched-Chain Amino Acid Oxidation and Incorporation into Proteins in Astrocytes and in Neurons in Primary Cultures 总被引:1,自引:1,他引:1
14CO2 production and incorporation of label into proteins from the labeled branched-chain amino acids, leucine, valine, and isoleucine, were determined in primary cultures of neurons and of undifferentiated and differentiated astrocytes from mouse cerebral cortex in the absence and presence of 3 mM ammonium chloride. Production of 14CO2 from [1-14C]leucine and [1-14C]valine was larger than 14CO2 production from [U-14C]leucine and [U-14C]valine in both astrocytes and neurons. In most cases more 14CO2 was produced in astrocytes than in neurons. Incorporation of labeled branched-chain amino acids into proteins varied with the cell type and with the amino acid. Addition of 3 mM ammonium chloride greatly suppressed 14CO2 production from [1-14C]-labeled branched chain amino acids but had little effect on 14CO2 production from [U-14C]-labeled branched-chain amino acids in astrocytes. Ammonium ion, at this concentration, suppressed the incorporation of label from all three branched-chain amino acids into proteins of astrocytes. In contrast, ammonium ion had very little effect on the metabolism (oxidation and incorporation into proteins) of these amino acids in neurons. The possible implications of these findings are discussed, especially regarding whether they signify variations in metabolic fluxes and/or in magnitudes of precursor pools. 相似文献
20.
Pawel Listwan Thomas C. Terwilliger Geoffrey S. Waldo 《Journal of structural and functional genomics》2009,10(1):47-55
Overproduction of soluble and stable proteins for functional and structural studies is a major bottleneck for structural genomics
programs and traditional biochemistry laboratories. Many high-payoff proteins that are important in various biological processes
are “difficult to handle” as protein reagents in their native form. We have recently made several advances in enabling biochemical
technologies for improving protein stability (), allowing stratagems for efficient protein domain trapping, solubility-improving mutations, and finding protein folding
partners. In particular split-GFP protein tags are a very powerful tool for detection of stable protein domains. Soluble,
stable proteins tagged with the 15 amino acid GFP fragment (amino acids 216–228) can be detected in vivo and in vitro using
the engineered GFP 1–10 “detector” fragment (amino acids 1–215). If the small tag is accessible, the detector fragment spontaneously
binds resulting in fluorescence. Here, we describe our current and on-going efforts to move this process from the bench (manual
sample manipulation) to an automated, high-throughput, liquid-handling platform. We discuss optimization and validation of
bacterial culture growth, lysis protocols, protein extraction, and assays of soluble and insoluble protein in multiple 96
well plate format. The optimized liquid-handling protocol can be used for rapid determination of the optimal, compact domains
from single ORFS, collections of ORFS, or cDNA libraries. 相似文献