• Title/Summary/Keyword: Biophysics

Search Result 555, Processing Time 0.023 seconds

The Alteration of $Ca^{2+}-activated\;K^+$ Channels in Coronary Arterial Smooth Muscle Cells Isolated from Isoproterenol-induced Cardiac Hypertrophy in Rabbit

  • Kim, Na-Ri;Han, Jin;Kim, Eui-Yong
    • The Korean Journal of Physiology and Pharmacology
    • /
    • v.5 no.2
    • /
    • pp.147-156
    • /
    • 2001
  • It has been proposed that $Ca^{2+}-activated$ K $(K_{Ca})$ channels play an essential role in vascular tone. The alterations of the properties of coronary $K_{Ca}$ channels have not been studied as a possible mechanism for impaired coronary reserve in cardiac hypertrophy. The present studies were carried out to determine the properties of coronary $K_{Ca}$ channels in normal and hypertrophied hearts. These channels were measured from rabbit coronary smooth muscle cells using a patch clamp technique. The main findings of the present study are as follows: (1) the unitary current amplitudes and the slope conductance of coronary $K_{Ca}$ channels were decreased without changes of the channel kinetics in isoproterenol-induced cardiac hypertrophy; (2) the sensitivity of coronary $K_{Ca}$ channels to the changes of intracellular concentration of $Ca^{2+}$ was reduced in isoproterenol-induced cardiac hypertrophy. From above results, we suggest for the first time that the alteration of $K_{Ca}$ channels are involved in impaired coronary reserve in isoproterenol-induced cardiac hypertrophy.

  • PDF

Effects of Nitric Oxide on Inhibitory Receptors of Rod Bipolar Cells of Rat Retina

  • Park, No-Gi;Bai, Sun-Ho;Jung, Chang-sub;Chun, Mynng-Hoon
    • The Korean Journal of Physiology and Pharmacology
    • /
    • v.9 no.6
    • /
    • pp.347-352
    • /
    • 2005
  • The effects of nitric oxide (NO) on inhibitory neurotransmitter receptors and some types of inhibitory receptors in dissociated rod bipolar cell (RBC) were investigated. In the whole cell voltage-clamping mode, the gamma-aminobutyric acid (GABA) activated current showed both sustained and transient components. GABA activated transient current was fully blocked by bicuculine, a $GABA_A$ receptor antagonist. The cis-4-aminocrotonic acid (CACA), a $GABA_C$ receptor agonist, evoked the sustained current that was not blocked by bicuculline (BIC). Glycine activated the transient current. These results indicate that the RBCs possess $GABA_A$, $GABA_C$, and glycine inhibitory receptors. Sodium nitroprusside (SNP), a NO analogue, reduced the currents activated by $GABA_A$ receptor only, however, did not reduce the currents activated by either $GABA_C$ or glycine receptors. This study signifies further that only NO depresses the fast inhibitory response activated by $GABA_A$ receptor in RBC. We, therefore, postulate that NO might depress the light-on/off transient inhibitory responses in RBCs in the rat retina.

Expression, Purification and NMR studies of SH3YL1 SH3 domain

  • Shrestha, Pravesh;Yun, Ji-Hye;Lee, Weon-Tae
    • Journal of the Korean Magnetic Resonance Society
    • /
    • v.14 no.2
    • /
    • pp.105-116
    • /
    • 2010
  • SH3YL1, a novel protein containing one Src homology 3 domain at the carboxyl terminus was first detected in mouse anagen skin cDNA. This protein had a significant homology with YHRO 16c/Ysc 84, the yeast Src homology 3 domain-containing protein. The sequence identity was remarkable at the carboxyl and amino-terminal Src homology 3 domain, suggesting that the novel protein is a mouse homolog of the yeast protein and thus was termed as SH3YL1. SH3YL1 is composed of two domains, a DUF500 at N-termini and a SH3 domain at C-termini. In our study we cloned the SH3 domain in bacterial expression system in Escherichia coli using pET32a vector with TEV protease cleavage site and purified as a monomer using affinity chromatography. The N-terminal poly-Histidine tag was cleaved with TEV protease and target protein was used for backbone studies. Our study showed that SH3 domain primarily consists of $\beta$-sheet which is in consistence with previous result performed on the truncated SH3 domain of SH3YL1.

Solution Structure of YKR049C, a Putative Redox Protein from Saccharomyces cerevisiae

  • Jung, Jin-Won;Yee, Adelinda;Wu, Bin;Arrowsmith, Cheryl H.;Lee, Weon-Tae
    • BMB Reports
    • /
    • v.38 no.5
    • /
    • pp.550-554
    • /
    • 2005
  • YKR049C is a mitochondrial protein in Saccharomyces cerevisiae that is conserved among yeast species, including Candida albicans. However, no biological function for YKR049C has been ascribed based on its primary sequence information. In the present study, NMR spectroscopy was used to determine the putative biological function of YKR049C based on its solution structure. YKR049C shows a well-defined thioredoxin fold with a unique insertion of helices between two $\beta$-strands. The central $\beta$-sheet divides the protein into two parts; a unique face and a conserved face. The 'unique face' is located between ${\beta}2$ and ${\beta}3$. Interestingly, the sequences most conserved among YKR049C families are found on this 'unique face', which incorporates L109 to E114. The side chains of these conserved residues interact with residues on the helical region with a stretch of hydrophobic surface. A putative active site composed by two short helices and a single Cys97 was also well observed. Our findings suggest that YKR049C is a redox protein with a thioredoxin fold containing a single active cysteine.

Conformational Lock and Dissociative Thermal Inactivation of Lentil Seedling Amine Oxidase

  • Moosavi-Nejad, S. Zahra;Moosavi-Movahedi, Ali-Akbar;Rezaei-Tavirani, Mostafa;Floris, Giovanni;Medda, Rosaria
    • BMB Reports
    • /
    • v.36 no.2
    • /
    • pp.167-172
    • /
    • 2003
  • The kinetics of thermal inactivation of copper-containing amine oxidase from lentil seedlings were studied in a 100 mM potassium phosphate buffer, pH 7, using putrescine as the substrate. The temperature range was between $47-60^{\circ}C$. The thermal inactivation curves were not linear at 52 and $57^{\circ}C$; three linear phases were shown. The first phase gave some information about the number of dimeric forms of the enzyme that were induced by the higher temperatures using the "conformational lock" pertaining theory to oligomeric enzyme. The "conformational lock" caused two additional dimeric forms of the enzyme when the temperature increased to $57^{\circ}C$. The second and third phases were interpreted according to a dissociative thermal inactivation model. These phases showed that lentil amine oxidase was reversibly-dissociated before the irreversible thermal inactivation. Although lentil amine oxidase is not a thermostable enzyme, its dimeric structure can form "conformational lock," conferring a structural tolerance to the enzyme against heat stress.

An ESR Study of Amino Acid and Protein Free Radicals in Solution. Part IV. An ESR study of Gamma-Irradiated Amino Acids in Frozen Aqueous Solutions.

  • Sun-Joo Hong;D. E. Holmes;L. H. Piette.
    • Journal of the Korean Chemical Society
    • /
    • v.15 no.5
    • /
    • pp.256-265
    • /
    • 1971
  • An ESR study has been made on free radicals produced in frozen aqueous solutions (ices) of glycine, DL-${\alpha}$-alanine, DL-serine, L-cysteine, DL-leucine and DL-isoleucine by gamma-irradiation at dry ice temperature. All free radicals induced were decayed concomitant to the successive annealing but the radical species which is believed to be dominant seems to be stable even near the melting point of the ice. These dominant species were found to be identical to those resulted from direct action of radiation in the solid at room temperature. Small but significant changes in the spectra of glycine and DL-${\alpha}$-alanine were observed by varying the microwave power. These results seem to support the view that the spectra obtained were composite consisting of more than two different resonances having different power saturation characteristics. The relative contribution of unidentified resonances to the composite spectra was greater for solutions of low concentration. These resonances are assumed to be induced by indirect effects, mainly hydrogen abstraction by radiation produced hydroxyl radicals and also C-N bond cleavage by hydrated electrons.

  • PDF

Microorganism lipid droplets and biofuel development

  • Liu, Yingmei;Zhang, Congyan;Shen, Xipeng;Zhang, Xuelin;Cichello, Simon;Guan, Hongbin;Liu, Pingsheng
    • BMB Reports
    • /
    • v.46 no.12
    • /
    • pp.575-581
    • /
    • 2013
  • Lipid droplet (LD) is a cellular organelle that stores neutral lipids as a source of energy and carbon. However, recent research has emerged that the organelle is involved in lipid synthesis, transportation, and metabolism, as well as mediating cellular protein storage and degradation. With the exception of multi-cellular organisms, some unicellular microorganisms have been observed to contain LDs. The organelle has been isolated and characterized from numerous organisms. Triacylglycerol (TAG) accumulation in LDs can be in excess of 50% of the dry weight in some microorganisms, and a maximum of 87% in some instances. These microorganisms include eukaryotes such as yeast and green algae as well as prokaryotes such as bacteria. Some organisms obtain carbon from $CO_2$ via photosynthesis, while the majority utilizes carbon from various types of biomass. Therefore, high TAG content generated by utilizing waste or cheap biomass, coupled with an efficient conversion rate, present these organisms as bio-tech 'factories' to produce biodiesel. This review summarizes LD research in these organisms and provides useful information for further LD biological research and microorganism biodiesel development.

Identification of novel Leishmania major antigens that elicit IgG2a response in resistant and susceptible mice

  • MOHAMMADI Mohammad Reza;ZEINALI Majid;ARDESTANI Sussan K.;KARIMINIA Amina
    • Parasites, Hosts and Diseases
    • /
    • v.44 no.1 s.137
    • /
    • pp.43-48
    • /
    • 2006
  • Experimental murine models with high, intermediate and low levels of genetically based susceptibility to Leishmania major infection reproduce almost entire spectrum of clinical manifestations of the human disease. There are increasing non-comparative studies on immune responses against isolated antigens of L. major in different murine strains. The aim of the present study was to find out whether there is an antigen that can induce protective immune response in resistant and susceptible murine strains. To do that, crude antigenic extract of procyclic and metacyclic promastigotes of L. major was prepared and subjected to SDS-PAGE electrophoresis. Western-blotting was used to search for antigen(s) capable of raising high antibody level of IgG2a versus IgG1 in the sera of both infected resistant and susceptible strains. Two novel antigens from metacyclic promastigotes of L. major (140 and 152 kDa) were potentially able to induce specific dominant IgG2a responses in BALB/c and C57BU6 mice. The 2 antigens also reacted with IgG antibody of cutaneous leishmaniasis patients. We confirm that 140 and 152 kDa proteins of L. major promastigotes are inducing IgG production in mice and humans.

Production of nitric oxide by murine macrophages induced by lipophosphoglycan of Leishmania major

  • KAVOOSI Gholamreza;ARDESTANI Sussan K.;KARIMINIA Amina;TAVAKOLI Zahra
    • Parasites, Hosts and Diseases
    • /
    • v.44 no.1 s.137
    • /
    • pp.35-41
    • /
    • 2006
  • Protozoan parasites of the genus Leishmania cause a number of important human diseases. One of the key determinants of parasite infectivity and survival is the surface glycoconjugate lipophosphoglycan (LPG). In addition, LPG is shown to be useful as a transmission blocking vaccine. Since culture supernatant of parasite promastigotes is a good source of LPG, we made attempts to characterize functions of the culture supernatant, and membrane LPG isolated from metacyclic promastigotes of Leishmania major. The purification scheme included anion-exchange chromatography, hydrophobic interaction chromatography and cold methanol precipitation. The purity of supernatant LPG (sLPG) and membrane LPG (mLPG) was determined by SOS-PAGE and thin layer chromatography. The effect of mLPG and sLPG on nitric oxide (NO) production by murine macrophages cell line (J77 4.1 A) was studied. Both sLPG and mLPG induced NO production in a dose dependent manner but sLPG induced significantly higher amount of NO than mLPG. Our results show that sLPG is able to promote NO production by murine macrophages.

Involvement of the Phospholipase C β1 Pathway in Desensitization of the Carbachol-activated Nonselective Cationic Current in Murine Gastric Myocytes

  • Kim, Byung Joo;So, Insuk;Kim, Ki Whan
    • Molecules and Cells
    • /
    • v.22 no.1
    • /
    • pp.65-69
    • /
    • 2006
  • In murine gastrointestinal myocytes muscarinic stimulation activates nonselective cation channels via a G-protein and $Ca^{2+}$-dependent pathway. We recorded inward cationic currents following application of carbachol ($I_{CCh}$) to murine gastric myocytes held at -60 mV, using the whole-cell patch-clamp method. The properties of the inward cationic currents were similar to those of the nonselective cation channels activated by muscarinic stimulation in other gastrointestinal smooth muscle cells. CCh-induced $I_{CCh}$ and spontaneous decay of $I_{CCh}$ (desensitization of $I_{CCh}$) occurred. Unlike the situation in guinea pig gastric myocytes, desensitization was not affected by varying $[EGTA]_i$. Pretreatment with the PLC inhibitor (U73122) blocked the activation of $I_{CCh}$, and desensitization of $I_{CCh}$ was attenuated in PLC ${\beta}_1$ knock-out mice. These results suggest that the desensitization of $I_{CCh}$ in murine gastric myocytes is not due to a pathway dependent on intracellular $Ca^{2+}$ but to the PLC ${\beta}_1$ pathway.