Barley α‐amylase/subtilisin inhibitor shows inhibitory activity against endogenous xylanase isozyme I of malted barley: A novel protein function

2020 ◽  
Vol 44 (6) ◽  
Author(s):  
Junyong Sun ◽  
Feng Xu ◽  
Jian Lu
Keyword(s):  
Inhibitory Activity ◽  
Protein Function ◽  
Subtilisin Inhibitor ◽  
Novel Protein

scholarly journals Arginine is essential for the α-amylase inhibitory activity of the α-amylase/subtilisin inhibitor (BASI) from barley seeds

1993 ◽  
Vol 293 (1) ◽  
pp. 151-155 ◽  
Author(s):  
J Abe ◽  
U Sidenius ◽  
B Svensson
Keyword(s):  
Inhibitory Activity ◽  
Enzyme Inhibitor ◽  
Three Dimensional ◽  
Order Rate Constant ◽  
Alpha Amylase ◽  
Three Dimensional Model ◽  
Pseudo First Order Reaction ◽  
Arginine Residues ◽  
Subtilisin Inhibitor ◽  
Target Enzyme

Treatment of barley alpha-amylase/subtilisin inhibitor (BASI) with reagents specific for arginine, histidine, methionine and tyrosine residues and amino and carboxyl groups indicates that an arginine residue(s) is essential for its action on the target enzyme barley alpha-amylase 2. Phenylglyoxal modified eight out of 12 arginine residues in BASI. Kinetic analysis shows that the inactivation of BASI follows a pseudo-first-order reaction and is due to reaction with one molecule of phenylglyoxal; the second-order rate constant is determined to be 2.95 M-1.min-1. At pH 8.0, BASI and barley alpha-amylase 2 form an inactive 1:1 complex. The Ki value of this association is 2.2 x 10(-10) M. The alpha-amylase protects four arginine residues and also the alpha-amylase inhibitory activity of BASI against phenylglyoxal. When BASI from the phenylglyoxal-modified target enzyme-inhibitor complex is isolated and subjected to a second treatment with phenylglyoxal, four additional arginine residues are modified, with concomitant loss of the inhibitory activity. These results are discussed in relation to a three-dimensional model of BASI based on the known structure of the corresponding inhibitor from wheat.

Rapid Creation of a Novel Protein Function by in Vitro Coevolution

2005 ◽  
Vol 348 (5) ◽  
pp. 1273-1282 ◽  
Author(s):  
Zhilei Chen ◽  
Huimin Zhao
Keyword(s):  
Protein Function ◽  
Novel Protein

scholarly journals Design to Data for mutants of β-glucosidase B from Paenibacillus polymyxa: M319C, T431I, and K337D

2019 ◽  
Author(s):  
Peishan Huang ◽  
Stephanie C. Contreras ◽  
Eliana Bloomfield ◽  
Kristine Schmitz ◽  
Augustine Arredondo ◽  
...  
Keyword(s):  
Protein Function ◽  
Kinetic Data ◽  
Prediction Accuracy ◽  
Paenibacillus Polymyxa ◽  
Computational Algorithms ◽  
Enzyme Design ◽  
Data Set ◽  
Computational Tools ◽  
Protein Functions ◽  
Novel Protein

ABSTRACTThe use of computational tools has become an increasingly popular tool for engineering protein function. While there are numerous examples of computational tools enabling the design of novel protein functions, there remains room for improvement in both prediction accuracy and success. To improve algorithms for functional and stability predictions, we have initiated the development of a data set designed to be used for training new computational algorithms for enzyme design. To date our dataset is composed of over 129 mutants with associated expression levels, kinetic data, and thermal stability for the enzyme β-glucosidase B (BglB) from Paenibacillus polymyxa. In this study, we introduced three new variants (M319C, T431I, and K337D) to our existing dataset with the goal of cultivating a larger dataset to train new design algorithms and more broadly explore structure-function relationships in BglB.

scholarly journals Control of Aptamer Function Using Radiofrequency Magnetic Field

2011 ◽  
Vol 2011 ◽  
pp. 1-6 ◽  
Author(s):  
Kenichi Taira ◽  
Koichi Abe ◽  
Takayuki Ishibasi ◽  
Katsuaki Sato ◽  
Kazunori Ikebukuro
Keyword(s):  
Magnetic Field ◽  
Control System ◽  
Gold Nanoparticle ◽  
Inhibitory Activity ◽  
Protein Function ◽  
Local Heating ◽  
Activity Assay ◽  
Complementary Dna ◽  
Dna Strand ◽  
Control Protein

Remote control of aptamer function has allowed us to control protein function in space and time. Here, we propose a novel control system for aptamer function by radiofrequency magnetic field- (RFMF-) induced local heating of a gold nanoparticle conjugated with an aptamer. In this study, we used a 31-mer thrombin-binding aptamer (TBA), which can inhibit thrombin activity, as a model aptamer. We evaluated the RFMF control of the inhibitory activity of a gold nanoparticle-conjugated TBA. To evaluate the effect of RFMF on enzymatic activity, we utilized a complementary DNA strand that maintains the broken structure during the activity assay. We observed a decrease in the inhibitory activity of TBA after RFMF irradiation. It indicates that RFMF is capable of controlling the TBA structure. Because RFMF allows noninvasive control of aptamer function, this strategy is expected to be novel way of controlling aptamer drug activity.

scholarly journals Accelerated Discovery of Novel Protein Function in Cultured Human Cells

2005 ◽  
Vol 4 (9) ◽  
pp. 1319-1327 ◽  
Author(s):  
Emily Hodges ◽  
Jenny Stjerndahl Redelius ◽  
Weilin Wu ◽  
Christer Höög
Keyword(s):  
Protein Function ◽  
Human Cells ◽  
Cultured Human Cells ◽  
Novel Protein

scholarly journals EirA Is a Novel Protein Essential for Intracellular Replication of Coxiella burnetii

2020 ◽  
Vol 88 (6) ◽  
Author(s):  
Miku Kuba ◽  
Nitika Neha ◽  
Patrice Newton ◽  
Yi Wei Lee ◽  
Vicki Bennett-Wood ◽  
...  
Keyword(s):  
Coxiella Burnetii ◽  
Protein Function ◽  
Galleria Mellonella ◽  
Q Fever ◽  
Axenic Culture ◽  
Febrile Illness ◽  
Intracellular Replication ◽  
Content Type ◽  
Novel Protein

ABSTRACT The zoonotic bacterial pathogen Coxiella burnetii is the causative agent of Q fever, a febrile illness which can cause a serious chronic infection. C. burnetii is a unique intracellular bacterium which replicates within host lysosome-derived vacuoles. The ability of C. burnetii to replicate within this normally hostile compartment is dependent on the activity of the Dot/Icm type 4B secretion system. In a previous study, a transposon mutagenesis screen suggested that the disruption of the gene encoding the novel protein CBU2072 rendered C. burnetii incapable of intracellular replication. This protein, subsequently named EirA (essential for intracellular replication A), is indispensable for intracellular replication and virulence, as demonstrated by infection of human cell lines and in vivo infection of Galleria mellonella. The putative N-terminal signal peptide is essential for protein function but is not required for localization of EirA to the bacterial inner membrane compartment and axenic culture supernatant. In the absence of EirA, C. burnetii remains viable but nonreplicative within the host phagolysosome, as coinfection with C. burnetii expressing native EirA rescues the replicative defect in the mutant strain. In addition, while the bacterial ultrastructure appears to be intact, there is an altered metabolic profile shift in the absence of EirA, suggesting that EirA may impact overall metabolism. Most strikingly, in the absence of EirA, Dot/Icm effector translocation was inhibited even when EirA-deficient C. burnetii replicated in the wild type (WT)-supported Coxiella containing vacuoles. EirA may therefore have a novel role in the control of Dot/Icm activity and represent an important new therapeutic target.

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