Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P06889 (Mol)
 630,302 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Prolonged myocardial stretch typically leads to hypertrophy of cardiomyocytes. As integrins are cellular receptors of stretch, we hypothesize that integrin stimulation induces cardiomyocyte hypertrophy. Integrins of neonatal rat cardiomyocytes (NRCMs) were stimulated with a peptide containing the Arg-Gly-Asp (RGD) sequence for 24 h. For comparison, alpha(1)-adrenergic stimulation by phenylephrine (PE) for 24 h was applied. Saline-treated NRCMs were used as control. The hypertrophic response was quantified by measuring cell surface area (CSA). Phosphorylation of NO-synthase-1 (NOS1) was assessed by immunocytochemistry. CSA was increased by 38% (IQR 31-44%) with RGD and by 68% (IQR 64-84%) with PE versus control (both P < 0.001). NOS-1 phosphorylation was increased by 61% with RGD and by 21% with PE versus control (both P < 0.01). A general NOS-inhibitor (L-NAME) inhibited RGD-induced hypertrophy completely, but had no significant effect on PE-induced hypertrophy. Administration of NO-donor to NRCMs co-incubated with RGD + L-NAME partly restored hypertrophy (to 62% of the hypertrophic effect of RGD alone), but had no effect if incubated with PE + L-NAME. Ryanodine and BAPTA-AM inhibited RGD-induced hypertrophy completely but not that induced by PE. Integrin stimulation of NRCMs by RGD leads to hypertrophy, likely by activation of NOS-1. Abrogation of RGD-induced hypertrophic response upon NOS-inhibition and rescue of this hypertrophic effect by NO-donor suggest that integrin stimulation-induced hypertrophy of NRCMs is NO-dependent.
Mol Cell Biochem 2009 Jan
PMID:Integrin stimulation-induced hypertrophy in neonatal rat cardiomyocytes is NO-dependent. 1869 Apr 13

Thermal stability is of great importance for industrial enzymes. Here we explored the thermal-stable mechanism of thermophilic nitrile hydratases (NHases) utilizing a molecular dynamic simulation. At a nanosecond timescale, profiles of root mean square fluctuation (RMSF) of two thermophilic NHases, 1UGQ and 1V29, under enhancing thermal stress were carried out at 300 K, 320 K, 350 K and 370 K, respectively. Results showed that the region A1 (211-231 aa) and A2 (305-316 aa) in 1UGQ, region B1 (186-192 aa) in 1V29, and most of terminal ends in both enzymes are hyper-sensitive. Salt-bridge analyses revealed that in one hand, salt-bridges contributed to maintaining the rigid structure and stable performance of the thermophilic 1UGQ and 1V29; in the other hand, salt-bridges involved in thermal sensitive regions are relatively weak and prone to be broken at elevated temperature, thereby cannot hold the stable conformation of the spatial neighborhood. In 1V29, region A1 was stabilized by a well-organized hook-hook like cluster with multiple salt-bridge interactions, region A2 was stabilized by two strong salt-bridge interactions of GLU52-ARG332 and GLU334-ARG332. In 1UGQ, the absence of a charged residue decreased its thermal sensitivity of region B1, and the formation of a small beta-sheet containing a stable salt-bridge in C-beta-terminal significantly enhanced its thermal stability. By radius of gyration calculation containing or eliminating the thermal sensitive regions, we quantified the contribution of thermal sensitive regions for thermal sensitivity of 1UGQ and 1V29. Consequently, we presented strategies to improve thermal stability of the industrialized mesophilic NHase by introducing stable salt-bridge interactions into its thermal sensitive regions.
J Mol Graph Model 2008 Nov
PMID:Insights into thermal stability of thermophilic nitrile hydratases by molecular dynamics simulation. 1894 44

Although the energetic balance of forces stabilizing proteins has been established qualitatively over the last decades, quantification of the energetic contribution of particular interactions still poses serious problems. The reasons are the strong cooperativity and the interdependence ofnoncovalent interactions. Salt bridges are a typical example. One expects that ionizable side chains frequently form ion pairs in innumerable crystal structures. Since electrostatic attraction between opposite charges is strong per se, salt bridges can intuitively be regarded as an important factor stabilizing the native structure. Is that really so? In this chapter we critically reassess the available methods to delineate the role ofelectrostatic interactions and salt bridges to protein stability, and discuss the progress and the obstacles in this endeavor. The basic problem is that formation of salt bridges depends on the ionization properties of the participating groups, which is significantly influenced by the protein environment. Furthermore, salt bridges experience thermal fluctuations, continuously break and re-form, and their lifespan in solution is governed by the flexibility of the protein. Finally, electrostatic interactions are long-range and might be significant in the unfolded state, thus seriously influencing the energetic profile. Elimination of salt bridges by protonation/deprotonation at extreme pH or by mutation provides only rough energetic estimates, since there is no way to account for the nonadditive response of the protein moiety. From what we know so far, the strength of electrostatic interactions is strongly context-dependent, yet it is unlikely that salt bridges are dominant factors governing protein stability. Nevertheless, proteins from thermophiles and hyperthermophiles exhibit more, and frequently networked, salt bridges than proteins from the mesophilic counterparts. Increasing the thermal (not the thermodynamic) stability of proteins by optimization of charge-charge interactions is a good example for an evolutionary solution utilizing physical factors.
Methods Mol Biol 2009
PMID:Defining the role of salt bridges in protein stability. 1915 86

Peptide side chain interactions were studied by molecular dynamics simulation using explicit solvent on a peptide with the sequence AAARAAAAEAAEAAAARA. Three different protonation states of the glutamic acid side chains were simulated for four 20 ns runs each, a total simulation time of 240 ns. Two different salt bridge geometries were observed and the preferred geometry was found to depend on Glu - Arg residue spacing. Stable charge clusters were also observed, particularly in the fully charged peptide. Salt bridges were selectively interrupted upon protonation, with concomitant changes in secondary structure. The fully charged peptide was highly helical between residues 9 and 13, although protonation increased helicity near the N-terminus. The contribution of salt bridges to helix stability therefore depends on both position and relative position of charged residues within a sequence.
J Mol Model 2009 Oct
PMID:Contribution of arginine-glutamate salt bridges to helix stability. 1926 93

We have previously demonstrated that alpha2-adrenoceptors regulate hypothalamic magnocellular oxytocinergic (OXY) neurons in Sprague Dawley rats. Here we investigated whether activation/inhibition of alpha2-adrenoceptors may similarly trigger/downregulate the activity of OXY neurons in control Long Evans (+/+) and permanently osmotically stressed Brattleboro (di/di) rats. The effect of alpha2-adrenoceptor agonist, xylazine (XYL) and alpha2-adrenoceptor antagonists, atipamezole (ATIP), and idazoxan (IDX) were evaluated in the supraoptic (SON) and paraventricular (PVN) hypothalamic nuclei. Saline (SAL, 0.1 ml/100 g), XYL (10 mg/kg), ATIP, (1 mg/kg), and IDX (10 mg/kg) and IDX or ATIP followed by XYL were applied intraperitoneally. Rats were sacrificed 90 min later and Fos/OXY co-labelings analyzed in microscope. In control +/+ rats no or few Fos/OXY co-labelings occurred in SON and PVN. XYL significantly increased Fos incidence in OXY neurons in both nuclei. ATIP significantly suppressed the effect of XYL in both nuclei and IDX only in SON. In di/di controls 81% of OXY neurons in SON and 44% in PVN revealed Fos presence and XYL did not further elevate Fos number in SON OXY neurons and slightly increased Fos number in PVN. ATIP or IDX only partially reduced Fos in SAL or XYL treated di/di rats. Our data indicate that: (1) XYL stimulation is not effective in di/di rats because of sustained upregulation of OXY neurons activity and (2) neither ATIP nor IDX reduced significantly the OXY activity in control di/di rats. These findings suggest that alpha2-adrenoceptors have only a limited impact in maintaining OXY cells activity upregulation in PVN and SON of di/di rats.
Cell Mol Neurobiol 2009 Sep
PMID:Alpha2-adrenergic impact on hypothalamic magnocellular oxytocinergic neurons in long evans and brattleboro rats: effects of agonist and antagonists. 1929 90

Folding of the Protein G B1 domain (PGB1) shifts with increasing salt concentration from a cooperative assembly of inherently unstructured subdomains to an assembly of partly pre-folded structures. The salt-dependence of pre-folding contributes to the stability minimum observed at physiological salt conditions. Our conclusions are based on a study in which the reconstitution of PGB1 from two fragments was studied as a function of salt concentrations and temperature using circular dichroism spectroscopy. Salt was found to induce an increase in beta-hairpin structure for the C-terminal fragment (residues 41 - 56), whereas no major salt effect on structure was observed for the isolated N-terminal fragment (residues 1 - 41). In line with the increasing evidence on the interrelation between fragment complementation and stability of the corresponding intact protein, we also find that salt effects on reconstitution can be predicted from salt dependence of the stability of the intact protein. Our data show that our variant (which has the mutations T2Q, N8D, N37D and reconstitutes in a manner similar to the wild type) displays the lowest equilibrium association constant around physiological salt concentration, with higher affinity observed both at lower and higher salt concentration. This corroborates the salt effects on the stability towards denaturation of the intact protein, for which the stability at physiological salt is lower compared to both lower and higher salt concentrations. Hence we conclude that reconstitution reports on molecular factors that govern the native states of proteins.
Int J Mol Sci 2009 Apr 08
PMID:Protein GB1 folding and assembly from structural elements. 1946 25

The protein kinase SOS2 (Salt Overly Sensitive 2) is essential for salt-stress signaling and tolerance in Arabidopsis. SOS2 is known to be activated by calcium-SOS3 and by phosphorylation at its activation loop. SOS2 is autophosphorylated in vitro, but the autophosphorylation site and its role in salt tolerance are not known. In this study, we identified an autophosphorylation site in SOS2 and analyzed its role in the responses of Arabidopsis to salt stress. Mass spectrometry analysis showed that Ser 228 of SOS2 is autophosphorylated. When this site was mutated to Ala, the autophosphorylation rate of SOS2 decreased. The substrate phosphorylation by the mutated SOS2 was also less than that by the wild-type SOS2. In contrast, changing Ser228 to Asp to mimic the autophosphorylation enhanced substrate phosphorylation by SOS2. Complementation tests in a sos2 mutant showed that the S228A but not the S228D mutation partially disrupted the function of SOS2 in salt tolerance. We also show that activation loop phosphorylation at Thr168 and autophosphorylation at Ser228 cannot substitute for each other, suggesting that both are required for salt tolerance. Our results indicate that Ser 228 of SOS2 is autophosphorylated and that this autophosphorylation is important for SOS2 function under salt stress.
Mol Plant 2009 Jan
PMID:An autophosphorylation site of the protein kinase SOS2 is important for salt tolerance in Arabidopsis. 1952 20

The lack of appropriate methods for storing intact and viable cells for the purpose of delayed DNA strand break analysis has hitherto limited the application of the Comet assay to in vitro or in vivo laboratory studies and restricted ecologically more relevant field-collected samples to sites in proximity to suitable laboratory facilities. In the present article, osmotically corrected cell culture media Hanks Balanced Salt Solution (HBSS) and Leibovitz Media (L-15) were assessed for their suitability as temporary storage media of blue mussel (Mytilus edulis) hemocytes. It was found that hemocytes maintained in either HBSS or L-15 could be stored for at least 7 days at 4 degrees C without any significant deterioration in cell viability (Trypan blue) or increase in DNA strand breaks, expressed as % tail DNA. This approach allows the acquisition and examination of samples from organisms exposed in situ at previously unsuitable remote sites, thereby greatly increasing the potential ecological relevance of Comet assay-derived genotoxicity data.
Environ Mol Mutagen 2010 Jan
PMID:Maintenance of bivalve hemocytes for the purpose of delayed DNA strand break assessment using the comet assay. 1959 4

Vacuolar H(+)-pyrophosphatase (V-PPase) expression increases in a number of abiotic stresses and is thought to play a role in adaptation to abiotic stresses. This paper reports on the regulation of six V-PPase genes in rice (Oryza sativa L.) coleoptiles under anoxia, using flood tolerant and intolerant cultivars to test our hypothesis. Quantitative PCR analysis showed that one vacuolar H(+)-pyrophosphatase (OVP3) was induced by anoxia, particularly in flood-tolerant rice. Regulation of OVP3 expression under anoxia was investigated by analysing putative OVP promoters. The putative OVP3 promoter contained more previously identified anoxia-inducible motifs than the putative promoters of the other five OVP genes. GUS activity in transgenic rice plants containing the OVP3 promoter region linked to the GUS reporter gene was induced only by anoxia. Salt and cold treatments had little effect on OVP3 promoter-driven GUS expression when compared to the anoxic treatment.
Plant Mol Biol 2010 Jan
PMID:Expression of vacuolar H+-pyrophosphatase (OVP3) is under control of an anoxia-inducible promoter in rice. 1976 43

Salt cress (Thellungiella halophila), a salt-tolerant relative of Arabidopsis, has turned to be an important model plant for studying abiotic stress tolerance. One binary bacterial artificial chromosome (BIBAC) library was constructed which represents the first plant-transformation-competent large-insert DNA library generated for Thellungiella halophila. The BIBAC library was constructed in BamHI site of binary vector pBIBAC2 by ligation of partial digested nuclear DNA of Thellungiella halophila. This library consists of 23,040 clones with an average insert size of 75 kb, and covers 4x Thellungiella halophila haploid genomes. BIBAC clones which contain inserts over 50 kb were selected and transformed into Arabidopsis for salt tolerant plant screening. One transgenic line was found to be more salt tolerant than wild type plants from the screen of 200 lines. It was demonstrated that the library contains candidates of stress tolerance genes and the approach is suitable for the transformation of stress susceptible plants for genetic improvement.
Plant Mol Biol 2010 Jan
PMID:A large insert Thellungiella halophila BIBAC library for genomics and identification of stress tolerance genes. 1978 33


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