Differential subcellular location and function of two isoenzymes of cardiac adenylate kinase

1981 ◽  
Vol 9 (6) ◽  
pp. 570-571
Author(s):  
ELAINE J. WALKER ◽  
JOCELYN W. DOW
Keyword(s):  
Adenylate Kinase ◽  
Subcellular Location ◽  
And Function

scholarly journals Chemical and physical properties of an arabinogalactan-peptide from wheat endosperm

1974 ◽  
Vol 139 (3) ◽  
pp. 535-545 ◽  
Author(s):  
G. B. Fincher ◽  
W. H. Sawyer ◽  
B. A. Stone
Keyword(s):  
Optical Rotation ◽  
Subcellular Location ◽  
Successive Treatment ◽  
Partial Acid Hydrolysis ◽  
Wheat Endosperm ◽  
Anomeric Configuration ◽  
Molecular Core ◽  
Tentative Model ◽  
And Function ◽  
Hydrolysis Of

1. An arabinogalactan-peptide from wheat endosperm was studied by using physicochemical techniques and some aspects of its chemical structure were determined. 2. The arabinogalactan-peptide is a non-associating, polydisperse macromolecule ([unk]=22000) which exhibits only minor non-ideal effects in aqueous solution. 3. Examination of the products of partial acid hydrolysis of the polysaccharide component showed that arabinose is present in the α-l-arabinofuranosyl configuration, and i.r.-absorption spectroscopy and optical-rotation studies suggest that the d-galactopyranose residues are linked by glycosidic linkages in the β-anomeric configuration. 4. The arabinogalactan is linked to a peptide which represents 8% (w/w) of the arabinogalactan-peptide and which may be present as a molecular core. Partial degradation of the polymer by successive treatment with oxalic acid and NaOH showed that the linkage between polysaccharide and peptide involves galactose and hydroxyproline residues and is glycosidic in nature. A tentative model is proposed for the structure of the wheat endosperm arabinogalactan-peptide. 5. The subcellular location and function of the arabinogalactan-peptide is discussed in relation to previous work with related molecules.

Red cell adenylate kinase deficiency: molecular study of 3 new mutations (118G>A, 190G>A, and GAC deletion) associated with hereditary nonspherocytic hemolytic anemia

Blood ◽  
2003 ◽  
Vol 102 (1) ◽  
pp. 353-356 ◽  
Author(s):  
Joan-Lluis Vives Corrons ◽  
Estefania Garcia ◽  
Joan J. Tusell ◽  
Kottayil I. Varughese ◽  
Carol West ◽  
...  
Keyword(s):  
Hemolytic Anemia ◽  
Adenylate Kinase ◽  
Cdna Sequence ◽  
Molecular Model ◽  
Molecular Study ◽  
Missense Mutations ◽  
White American ◽  
And Function ◽  
Enzyme Structure And Function ◽  
New Mutations

Abstract We report here 2 patients with chronic nonspherocytic hemolytic anemia (CNSHA) and severe red blood cell (RBC) adenylate kinase (AK) deficiency. One of these patients, a boy of Spanish origin, exhibited a neonatal icterus and splenomegaly and required blood transfusions until the age of 2 years. The other patient was a white, American infant born to parents who were first cousins; he also presented with neonatal icterus and anemia. In neither case was psychomotor impairment observed. The first patient was found to be a compound heterozygote for 2 different missense mutations, 118G>A(Gly40Arg) and 190G>A(Gly64Arg) (cDNA sequence first described by Matsuura et al, 1989). The second patient was homozygous for an in-frame deletion (GAC) from nucleotide (nt) 498 to 500 or nt 501 to 503 of the cDNA sequence, predicting deletion of either aspartic acid (Asp) 140 or 141. The crystal structure of porcine cytosolic AK was used as a molecular model to investigate how these mutations may affect enzyme structure and function. (Blood. 2003;102:353-356)

scholarly journals Mitochondria-specific photoactivation to monitor local sphingosine metabolism and function

eLife ◽  
2018 ◽  
Vol 7 ◽  
Author(s):  
Suihan Feng ◽  
Takeshi Harayama ◽  
Sylvie Montessuit ◽  
Fabrice PA David ◽  
Nicolas Winssinger ◽  
...  
Keyword(s):  
Time Course ◽  
Random Distribution ◽  
Direct Evidence ◽  
Subcellular Location ◽  
Sphingolipid Metabolism ◽  
Powerful Approach ◽  
Sphingosine 1 Phosphate ◽  
Sphingosine Kinases ◽  
And Function ◽  
Caged Molecules

Photoactivation ('uncaging’) is a powerful approach for releasing bioactive small-molecules in living cells. Current uncaging methods are limited by the random distribution of caged molecules within cells. We have developed a mitochondria-specific photoactivation method, which permitted us to release free sphingosine inside mitochondria and thereafter monitor local sphingosine metabolism by lipidomics. Our results indicate that sphingosine was quickly phosphorylated into sphingosine 1-phosphate (S1P) driven by sphingosine kinases. In time-course studies, the mitochondria-specific uncaged sphingosine demonstrated distinct metabolic patterns compared to globally-released sphingosine, and did not induce calcium spikes. Our data provide direct evidence that sphingolipid metabolism and signaling are highly dependent on the subcellular location and opens up new possibilities to study the effects of lipid localization on signaling and metabolic fate.

scholarly journals A new method for post-translationally labeling proteins in live cells for fluorescence imaging and tracking

2017 ◽  
Vol 30 (12) ◽  
pp. 771-780 ◽  
Author(s):  
M Hinrichsen ◽  
M Lenz ◽  
J M Edwards ◽  
O K Miller ◽  
S G J Mochrie ◽  
...  
Keyword(s):  
Fluorescent Protein ◽  
Single Cells ◽  
Covalent Bond ◽  
Subcellular Location ◽  
Turnover Time ◽  
Target Protein ◽  
Live Cells ◽  
General Applicability ◽  
Novel Approach ◽  
And Function

AbstractWe present a novel method to fluorescently label proteins, post-translationally, within live Saccharomycescerevisiae. The premise underlying this work is that fluorescent protein (FP) tags are less disruptive to normal processing and function when they are attached post-translationally, because target proteins are allowed to fold properly and reach their final subcellular location before being labeled. We accomplish this post-translational labeling by expressing the target protein fused to a short peptide tag (SpyTag), which is then covalently labeled in situ by controlled expression of an open isopeptide domain (SpyoIPD, a more stable derivative of the SpyCatcher protein) fused to an FP. The formation of a covalent bond between SpyTag and SpyoIPD attaches the FP to the target protein. We demonstrate the general applicability of this strategy by labeling several yeast proteins. Importantly, we show that labeling the membrane protein Pma1 in this manner avoids the mislocalization and growth impairment that occur when Pma1 is genetically fused to an FP. We also demonstrate that this strategy enables a novel approach to spatiotemporal tracking in single cells and we develop a Bayesian analysis to determine the protein’s turnover time from such data.

scholarly journals The Subcellular Location of Ovalbumin in Plasmodium berghei Blood Stages Influences the Magnitude of T-Cell Responses

2014 ◽  
Vol 82 (11) ◽  
pp. 4654-4665 ◽  
Author(s):  
Jing-Wen Lin ◽  
Tovah N. Shaw ◽  
Takeshi Annoura ◽  
Aurélie Fougère ◽  
Pascale Bouchier ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Plasmodium Berghei ◽  
Parasitophorous Vacuole ◽  
T Cell Responses ◽  
Subcellular Location ◽  
Expression Level ◽  
Content Type ◽  
Cell Responses ◽  
And Function

ABSTRACTModel antigens are frequently introduced into pathogens to study determinants that influence T-cell responses to infections. To address whether an antigen's subcellular location influences the nature and magnitude of antigen-specific T-cell responses, we generatedPlasmodium bergheiparasites expressing the model antigen ovalbumin (OVA) either in the parasite cytoplasm or on the parasitophorous vacuole membrane (PVM). For cytosolic expression, OVA alone or conjugated to mCherry was expressed from a strong constitutive promoter (OVAhsp70orOVA::mCherryhsp70); for PVM expression, OVA was fused to HEP17/EXP1 (OVA::Hep17hep17). Unexpectedly, OVA expression inOVAhsp70parasites was very low, but when OVA was fused to mCherry (OVA::mCherryhsp70), it was highly expressed. OVA expression inOVA::Hep17hep17parasites was strong but significantly less than that inOVA::mCherryhsp70parasites. These transgenic parasites were used to examine the effects of antigen subcellular location and expression level on the development of T-cell responses during blood-stage infections. While all OVA-expressing parasites induced activation and proliferation of OVA-specific CD8+T cells (OT-I) and CD4+T cells (OT-II), the level of activation varied:OVA::Hep17hep17parasites induced significantly stronger splenic and intracerebral OT-I and OT-II responses than those ofOVA::mCherryhsp70parasites, butOVA::mCherryhsp70parasites promoted stronger OT-I and OT-II responses than those ofOVAhsp70parasites. Despite lower OVA expression levels,OVA::Hep17hep17parasites induced stronger T-cell responses than those ofOVA::mCherryhsp70parasites. These results indicate that unconjugated cytosolic OVA is not stably expressed inPlasmodiumparasites and, importantly, that its cellular location and expression level influence both the induction and magnitude of parasite-specific T-cell responses. These parasites represent useful tools for studying the development and function of antigen-specific T-cell responses during malaria infection.

scholarly journals A nanobody-based molecular toolkit provides new mechanistic insight into clathrin-coat initiation

eLife ◽  
2019 ◽  
Vol 8 ◽  
Author(s):  
Linton M Traub
Keyword(s):  
Decisive Role ◽  
Subcellular Location ◽  
Combined Approach ◽  
Mechanistic Insight ◽  
Early Endosomes ◽  
The Stability ◽  
And Function ◽  
Insight Into ◽  
Complementarity Determining Region

Besides AP-2 and clathrin triskelia, clathrin coat inception depends on a group of early-arriving proteins including Fcho1/2 and Eps15/R. Using genome-edited cells, we described the role of the unstructured Fcho linker in stable AP-2 membrane deposition. Here, expanding this strategy in combination with a new set of llama nanobodies against EPS15 shows an FCHO1/2–EPS15/R partnership plays a decisive role in coat initiation. A nanobody containing an Asn-Pro-Phe peptide within the complementarity-determining region 3 loop is a function-blocking pseudoligand for tandem EPS15/R EH domains. Yet, in living cells, EH domains gathered at clathrin-coated structures are poorly accessible, indicating residence by endogenous NPF-bearing partners. Forcibly sequestering cytosolic EPS15 in genome-edited cells with nanobodies tethered to early endosomes or mitochondria changes the subcellular location and availability of EPS15. This combined approach has strong effects on clathrin coat structure and function by dictating the stability of AP-2 assemblies at the plasma membrane.

scholarly journals Extracellular loops matter – subcellular location and function of the lysine transporter Lyp1 from Saccharomyces cerevisiae

FEBS Journal ◽  
2020 ◽  
Vol 287 (20) ◽  
pp. 4401-4414 ◽  
Author(s):  
Joury S. van‘t Klooster ◽  
Frans Bianchi ◽  
Ruben B. Doorn ◽  
Mirco Lorenzon ◽  
Jarnick H. Lusseveld ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Subcellular Location ◽  
Extracellular Loops ◽  
And Function

scholarly journals Purine-Metabolizing Ectoenzymes Control IL-8 Production in Human Colon HT-29 Cells

2014 ◽  
Vol 2014 ◽  
pp. 1-10 ◽  
Author(s):  
Fariborz Bahrami ◽  
Filip Kukulski ◽  
Joanna Lecka ◽  
Alain Tremblay ◽  
Julie Pelletier ◽  
...  
Keyword(s):  
Adenylate Kinase ◽  
Adenine Nucleotide ◽  
Inflammatory Diseases ◽  
Cancer Cell Line ◽  
Human Colon ◽  
Nucleotide Metabolism ◽  
Poly I:C ◽  
Intestinal Epithelial ◽  
And Function ◽  
Ht 29

Interleukin-8 (IL-8) plays key roles in both chronic inflammatory diseases and tumor modulation. We previously observed that IL-8 secretion and function can be modulated by nucleotide (P2) receptors. Here we investigated whether IL-8 release by intestinal epithelial HT-29 cells, a cancer cell line, is modulated by extracellular nucleotide metabolism. We first identified that HT-29 cells regulated adenosine and adenine nucleotide concentration at their surface by the expression of the ectoenzymes NTPDase2, ecto-5′-nucleotidase, and adenylate kinase. The expression of the ectoenzymes was evaluated by RT-PCR, qPCR, and immunoblotting, and their activity was analyzed by RP-HPLC of the products and by detection ofPiproduced from the hydrolysis of ATP, ADP, and AMP. In response to poly (I:C), with or without ATP and/or ADP, HT-29 cells released IL-8 and this secretion was modulated by the presence of NTPDase2 and adenylate kinase. Taken together, these results demonstrate the presence of 3 ectoenzymes at the surface of HT-29 cells that control nucleotide levels and adenosine production (NTPDase2, ecto-5′-nucleotidase and adenylate kinase) and that P2 receptor-mediated signaling controls IL-8 release in HT-29 cells which is modulated by the presence of NTPDase2 and adenylate kinase.

scholarly journals Axonal and dendritic synaptotagmin isoforms revealed by a pHluorin-syt functional screen

2012 ◽  
Vol 23 (9) ◽  
pp. 1715-1727 ◽  
Author(s):  
Camin Dean ◽  
F. Mark Dunning ◽  
Huisheng Liu ◽  
Ewa Bomba-Warczak ◽  
Henrik Martens ◽  
...  
Keyword(s):  
Membrane Fusion ◽  
Synaptic Vesicles ◽  
Ph Sensor ◽  
Subcellular Location ◽  
Neuronal Function ◽  
Specific Antibodies ◽  
Functional Screen ◽  
And Function ◽  
The Brain

The synaptotagmins (syts) are a family of molecules that regulate membrane fusion. There are 17 mammalian syt isoforms, most of which are expressed in the brain. However, little is known regarding the subcellular location and function of the majority of these syts in neurons, largely due to a lack of isoform-specific antibodies. Here we generated pHluorin-syt constructs harboring a luminal domain pH sensor, which reports localization, pH of organelles to which syts are targeted, and the kinetics and sites of exocytosis and endocytosis. Of interest, only syt-1 and 2 are targeted to synaptic vesicles, whereas other isoforms selectively recycle in dendrites (syt-3 and 11), axons (syt-5, 7, 10, and 17), or both axons and dendrites (syt-4, 6, 9, and 12), where they undergo exocytosis and endocytosis with distinctive kinetics. Hence most syt isoforms localize to distinct secretory organelles in both axons and dendrites and may regulate neuropeptide/neurotrophin release to modulate neuronal function.

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