scholarly journals Crystal structure and tartrate inhibition of Legionella pneumophila histidine acid phosphatase

2015 ◽  
Vol 585 ◽  
pp. 32-38 ◽  
Author(s):  
Richa Dhatwalia ◽  
Harkewal Singh ◽  
Thomas J. Reilly ◽  
John J. Tanner
Keyword(s):  
Crystal Structure ◽  
Acid Phosphatase ◽  
Legionella Pneumophila ◽  
Histidine Acid Phosphatase

scholarly journals N-Glycosylation Improves the Pepsin Resistance of Histidine Acid Phosphatase Phytases by Enhancing Their Stability at Acidic pHs and Reducing Pepsin's Accessibility to Its Cleavage Sites

2015 ◽  
Vol 82 (4) ◽  
pp. 1004-1014 ◽  
Author(s):  
Canfang Niu ◽  
Huiying Luo ◽  
Pengjun Shi ◽  
Huoqing Huang ◽  
Yaru Wang ◽  
...  
Keyword(s):  
Acid Phosphatase ◽  
Biochemical Characterization ◽  
Animal Feed ◽  
Glycosylation Site ◽  
Site Directed Mutagenesis ◽  
Acidic Ph ◽  
Cleavage Sites ◽  
Ph Optimum ◽  
Content Type ◽  
Histidine Acid Phosphatase

ABSTRACTN-Glycosylation can modulate enzyme structure and function. In this study, we identified two pepsin-resistant histidine acid phosphatase (HAP) phytases fromYersinia kristensenii(YkAPPA) andYersinia rohdei(YrAPPA), each having anN-glycosylation motif, and one pepsin-sensitive HAP phytase fromYersinia enterocolitica(YeAPPA) that lacked anN-glycosylation site. Site-directed mutagenesis was employed to construct mutants by altering theN-glycosylation status of each enzyme, and the mutant and wild-type enzymes were expressed inPichia pastorisfor biochemical characterization. Compared with those of theN-glycosylation site deletion mutants andN-deglycosylated enzymes, allN-glycosylated counterparts exhibited enhanced pepsin resistance. Introduction of theN-glycosylation site into YeAPPA as YkAPPA and YrAPPA conferred pepsin resistance, shifted the pH optimum (0.5 and 1.5 pH units downward, respectively) and improved stability at acidic pH (83.2 and 98.8% residual activities at pH 2.0 for 1 h). Replacing the pepsin cleavage sites L197 and L396 in the immediate vicinity of theN-glycosylation motifs of YkAPPA and YrAPPA with V promoted their resistance to pepsin digestion when produced inEscherichia colibut had no effect on the pepsin resistance ofN-glycosylated enzymes produced inP. pastoris. Thus,N-glycosylation may improve pepsin resistance by enhancing the stability at acidic pH and reducing pepsin's accessibility to peptic cleavage sites. This study provides a strategy, namely, the manipulation ofN-glycosylation, for improvement of phytase properties for use in animal feed.

scholarly journals Protein polyglutamylation catalyzed by the bacterial calmodulin-dependent pseudokinase SidJ

eLife ◽  
2019 ◽  
Vol 8 ◽  
Author(s):  
Alan Sulpizio ◽  
Marena E Minelli ◽  
Min Wan ◽  
Paul D Burrowes ◽  
Xiaochun Wu ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Mass Spectrometry ◽  
Ubiquitin Ligase ◽  
Legionella Pneumophila ◽  
Kinase Activity ◽  
Effector Protein ◽  
Dependent Manner ◽  
Biochemical Analyses ◽  
Human And Mouse

Pseudokinases are considered to be the inactive counterparts of conventional protein kinases and comprise approximately 10% of the human and mouse kinomes. Here, we report the crystal structure of the Legionella pneumophila effector protein, SidJ, in complex with the eukaryotic Ca2+-binding regulator, calmodulin (CaM). The structure reveals that SidJ contains a protein kinase-like fold domain, which retains a majority of the characteristic kinase catalytic motifs. However, SidJ fails to demonstrate kinase activity. Instead, mass spectrometry and in vitro biochemical analyses demonstrate that SidJ modifies another Legionella effector SdeA, an unconventional phosphoribosyl ubiquitin ligase, by adding glutamate molecules to a specific residue of SdeA in a CaM-dependent manner. Furthermore, we show that SidJ-mediated polyglutamylation suppresses the ADP-ribosylation activity. Our work further implies that some pseudokinases may possess ATP-dependent activities other than conventional phosphorylation.

scholarly journals Crystal structure of a lipin/Pah phosphatidic acid phosphatase

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Valerie I. Khayyo ◽  
Reece M. Hoffmann ◽  
Huan Wang ◽  
Justin A. Bell ◽  
John E. Burke ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Acid Phosphatase ◽  
Phosphatidic Acid ◽  
Phosphatidic Acid Phosphatase

Crystal structure of a mammalian purple acid phosphatase 1 1Edited by R. Huber

1999 ◽  
Vol 290 (1) ◽  
pp. 201-211 ◽  
Author(s):  
Jonas Uppenberg ◽  
Fredrik Lindqvist ◽  
Carina Svensson ◽  
Barbro Ek-Rylander ◽  
Göran Andersson
Keyword(s):  
Crystal Structure ◽  
Acid Phosphatase ◽  
Purple Acid Phosphatase

scholarly journals The Legionnaires' disease bacterium (Legionella pneumophila) inhibits phagosome-lysosome fusion in human monocytes.

1983 ◽  
Vol 158 (6) ◽  
pp. 2108-2126 ◽  
Author(s):  
M A Horwitz
Keyword(s):  
Acid Phosphatase ◽  
Legionella Pneumophila ◽  
Thorium Dioxide ◽  
Live Bacteria ◽  
Phosphatase Cytochemistry ◽  
Legionnaires Disease ◽  
Intracellular Multiplication ◽  
Erythromycin A ◽  
Secondary Lysosomes ◽  
Bacterial Protein Synthesis

The interactions between the L. pneumophila phagosome and monocyte lysosomes were investigated by prelabeling the lysosomes with thorium dioxide, an electron-opaque colloidal marker, and by acid phosphatase cytochemistry. Phagosomes containing live L. pneumophila did not fuse with secondary lysosomes at 1 h after entry into monocytes or at 4 or 8 h after entry by which time the ribosome-lined L. pneumophila replicative vacuole had formed. In contrast, the majority of phagosomes containing formalin-killed L. pneumophila, live Streptococcus pneumoniae, and live Escherichia coli had fused with secondary lysosomes by 1 h after entry into monocytes. Erythromycin, a potent inhibitor of bacterial protein synthesis, at a concentration that completely inhibits L. pneumophila intracellular multiplication, had no influence on fusion of L. pneumophila phagosomes with secondary lysosomes. However, coating live L. pneumophila with antibody or with antibody and complement partially overcame the inhibition of fusion. Also activating the monocytes promoted fusion of a small proportion of phagosomes containing live L. pneumophila with secondary lysosomes. Acid phosphatase cytochemistry revealed that phagosomes containing live L. pneumophila did not fuse with either primary or secondary lysosomes. In contrast to phagosomes containing live bacteria, the majority of phagosomes containing formalin-killed L. pneumophila were fused with lysosomes by acid phosphatase cytochemistry. The capacity of L. pneumophila to inhibit phagosome-lysosome fusion may be a critical mechanism by which the bacterium resists monocyte microbicidal effects.

scholarly journals Crystal structure of lpg1832, a VirK family protein from Legionella pneumophila , reveals a novel fold for bacterial VirK proteins

FEBS Letters ◽  
2017 ◽  
Vol 591 (18) ◽  
pp. 2929-2935 ◽  
Author(s):  
Nannan Zhang ◽  
Shiyan Yin ◽  
Shan Liu ◽  
Aihong Sun ◽  
Mingxue Zhou ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Legionella Pneumophila ◽  
Family Protein

scholarly journals The crystal structure of LidA, a translocated substrate of the Legionella pneumophila type IV secretion system

2013 ◽  
Vol 4 (12) ◽  
pp. 897-900 ◽  
Author(s):  
Geng Meng ◽  
Xiaojing An ◽  
Sheng Ye ◽  
Yong Liu ◽  
Wenzhuang Zhu ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Legionella Pneumophila ◽  
Secretion System ◽  
Type Iv Secretion ◽  
Type Iv Secretion System ◽  
Type Iv

scholarly journals Crystallization and preliminary crystallographic analysis of the major acid phosphatase fromLegionella pneumophila

2015 ◽  
Vol 71 (6) ◽  
pp. 779-783 ◽  
Author(s):  
Dan Zhou ◽  
Yang Pan ◽  
Xiaofang Chen ◽  
Nannan Zhang ◽  
Honghua Ge
Keyword(s):  
Acid Phosphatase ◽  
Unit Cell ◽  
Diffusion Method ◽  
Phosphoryl Group ◽  
Sequence Motif ◽  
Unit Cell Parameters ◽  
Space Group ◽  
Cell Parameters ◽  
Histidine Acid Phosphatase ◽  
Major Acid

The major acid phosphatase fromLegionella pneumophila(LpMAP) belongs to the histidine acid phosphatase superfamily. It contains the characteristic histidine acid phosphatase (HAP) sequence motif RHGXRXP responsible for the hydrolysis of a phosphoryl group from phosphate monoesters under acidic conditions. Here, the crystallization and preliminary X-ray analysis of crystals ofLpMAP in the apo form and in complex with L-(+)-tartrate are described. By using the hanging-drop vapour-diffusion method, apoLpMAP andLpMAP–tartrate were crystallized in space groupP21, with unit-cell parametersa= 91.50,b= 56.48,c= 146.35 Å, β = 110.01°, and in space groupP1, with unit-cell parametersa= 55.51,b= 73.51 ,c= 98.78 Å, α = 78.82, β = 77.65, γ = 67.73°, respectively. Diffraction data were collected at 100 K and the phases were determined using the molecular-replacement method.

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