scholarly journals Cysteine proteases of human hookworm Necator Americanus as virulence factors and implications for drug design with anti-heparin and heparin analogs: A bioinformatics study

2017 ◽  
Author(s):  
Arpita Banerjee ◽  
Kevin Widmer ◽  
Conor R Caffrey ◽  
Ruben A. Abagyan
Keyword(s):  
Cathepsin B ◽  
Cysteine Proteases ◽  
Cysteine Residue ◽  
Deficiency Anemia ◽  
Pathogenic Potential ◽  
Necator Americanus ◽  
Active Site Cysteine ◽  
Bioinformatics Study ◽  
Hemoglobin Degradation ◽  
Active Site Cysteine Residue

AbstractHuman hookworm Necator Americanus (NA) causes iron deficiency anemia, as the parasite ingests blood from the gastrointestinal tract of its human host. This bioinformatics-based study focuses on eight of the cathepsin B-like cysteine proteases (CPs) of the worm to explore their pathogenic potential. CP1 - CP6, which harbored the active site cysteine residue for enzymatic activity, were relevantly observed to have N-terminal signal peptide for extracellular localization. The secretory CPs could be releasing indigenous worm heparin at the host-pathogen interface for anticoagulation purposes. CP2 and CP3 showed a novel hemoglobinase motif that could be a prerequisite for hemoglobin degradation. CP1 and CP6 shared similar enzymatic-pocket features with cathepsin B and cruzain that cleave high molecular weight kininogen for blood-thinning activity. CP1, CP2, CP3, CP5 and CP6 were predicted to bind heparin, at their C terminal domain, like human cathepsin B and cruzain non-covalently bind heparin to enhance their activity. NA CPs’ action in concert with heparin, have implications for anti-heparin and heparin analog design against hookworm infection.

scholarly journals Evidence for inactivation of cysteine proteases by reactive carbonyls via glycation of active site thiols

2006 ◽  
Vol 398 (2) ◽  
pp. 197-206 ◽  
Author(s):  
Jingmin Zeng ◽  
Rachael A. Dunlop ◽  
Kenneth J. Rodgers ◽  
Michael J. Davies
Keyword(s):  
Active Site ◽  
Cysteine Proteases ◽  
Cysteine Residue ◽  
Dependent Manner ◽  
Cross Link ◽  
Concentration Dependent Manner ◽  
Active Site Cysteine ◽  
Reactive Aldehydes ◽  
Modified Proteins ◽  
Active Site Cysteine Residue

Hyperglycaemia, triose phosphate decomposition and oxidation reactions generate reactive aldehydes in vivo. These compounds react non-enzymatically with protein side chains and N-terminal amino groups to give adducts and cross-links, and hence modified proteins. Previous studies have shown that free or protein-bound carbonyls inactivate glyceraldehyde-3-phosphate dehydrogenase with concomitant loss of thiol groups [Morgan, Dean and Davies (2002) Arch. Biochem. Biophys. 403, 259–269]. It was therefore hypothesized that modification of lysosomal cysteine proteases (and the structurally related enzyme papain) by free and protein-bound carbonyls may modulate the activity of these components of the cellular proteolytic machinery responsible for the removal of modified proteins and thereby contribute to a decreased removal of modified proteins from cells. It is shown that MGX (methylglyoxal), GO (glyoxal) and glycolaldehyde, but not hydroxyacetone and glucose, inhibit catB (cathepsin B), catL (cathepsin L) and catS (cathepsin S) activity in macrophage cell lysates, in a concentration-dependent manner. Protein-bound carbonyls produced similar inhibition with both cell lysates and intact macrophage cells. Inhibition was also observed with papain, with this paralleled by loss of the active site cysteine residue and formation of the adduct species S-carboxymethylcysteine, from GO, in a concentration-dependent manner. Inhibition of autolysis of papain by MGX, along with cross-link formation, was detected by SDS/PAGE. Treatment of papain and catS with the dialdehyde o-phthalaldehyde resulted in enzyme inactivation and an intra-molecular active site cysteine–lysine cross-link. These results demonstrate that reactive aldehydes inhibit cysteine proteases by modification of the active site cysteine residue. This process may contribute to the accumulation of modified proteins in tissues of people with diabetes and age-related pathologies, including atherosclerosis, cataract and Alzheimer's disease.

scholarly journals The location of the active-site histidine residue in the primary sequence of papain

1968 ◽  
Vol 108 (5) ◽  
pp. 861-866 ◽  
Author(s):  
S. S. Husain ◽  
G. Lowe
Keyword(s):  
Amino Acid ◽  
Sodium Borohydride ◽  
Active Site ◽  
Iodoacetic Acid ◽  
Cysteine Residue ◽  
Histidine Residue ◽  
Primary Sequence ◽  
Active Site Cysteine ◽  
Active Site Histidine ◽  
Active Site Cysteine Residue

Papain that had been irreversibly inhibited with 1,3-dibromo[2−14C]acetone was reduced with sodium borohydride and carboxymethylated with iodoacetic acid. After digestion with trypsin and α-chymotrypsin the radioactive peptides were purified chromatographically. Their amino acid composition indicated that cysteine-25 and histidine-106 were cross-linked. Since cysteine-25 is known to be the active-site cysteine residue, histidine-106 must be the active-site histidine residue.

scholarly journals Analysis of the Active Site Cysteine Residue of the Sacrificial Sulfur Insertase LarE from Lactobacillus plantarum

Biochemistry ◽  
2018 ◽  
Vol 57 (38) ◽  
pp. 5513-5523 ◽  
Author(s):  
Matthias Fellner ◽  
Joel A. Rankin ◽  
Benoît Desguin ◽  
Jian Hu ◽  
Robert P. Hausinger
Keyword(s):  
Lactobacillus Plantarum ◽  
Active Site ◽  
Cysteine Residue ◽  
Active Site Cysteine ◽  
Active Site Cysteine Residue

OnBlastocystissecreted cysteine proteases: a legumain-activated cathepsin B increases paracellular permeability of intestinal Caco-2 cell monolayers

Parasitology ◽  
2016 ◽  
Vol 143 (13) ◽  
pp. 1713-1722 ◽  
Author(s):  
C. NOURRISSON ◽  
I. WAWRZYNIAK ◽  
A. CIAN ◽  
V. LIVRELLI ◽  
E. VISCOGLIOSI ◽  
...  
Keyword(s):  
Cathepsin B ◽  
Proteolytic Enzymes ◽  
Cell Monolayer ◽  
Cysteine Proteases ◽  
Cell Permeability ◽  
Whole Genome Sequence ◽  
Intestinal Cell ◽  
Pathogenic Potential ◽  
Cell Monolayers ◽  
Culture Supernatants

SUMMARYBlastocystisspp. pathogenic potential remains unclear as these anaerobic parasitic protozoa are frequently isolated from stools of both symptomatic and asymptomatic subjects.In silicoanalysis of the whole genome sequence ofBlastocystissubtype 7 revealed the presence of numerous proteolytic enzymes including cysteine proteases predicted to be secreted. To assess the potential impact of proteases on intestinal cells and gut function, we focused our study on two cysteine proteases, a legumain and a cathepsin B, which were previously identified inBlastocystissubtype 7 culture supernatants. Both cysteine proteases were produced as active recombinant proteins. Activation of the recombinant legumain was shown to be autocatalytic and triggered by acidic pH, whereas proteolytic activity of the recombinant cathepsin B was only recorded after co-incubation with the legumain. We then measured the diffusion of 4-kDa FITC-labelled dextran across Caco-2 cell monolayers following exposition to eitherBlastocystisculture supernatants or each recombinant protease. BothBlastocystisculture supernatants and recombinant activated cathepsin B induced an increase of Caco-2 cell monolayer permeability, and this effect was significantly inhibited by E-64, a specific cysteine protease inhibitor. Our results suggest that cathepsin B might play a role in pathogenesis ofBlastocystisby increasing intestinal cell permeability.

scholarly journals Inactivation of Ca2+/calmodulin-dependent protein kinase I byS-glutathionylation of the active-site cysteine residue

FEBS Letters ◽  
2010 ◽  
Vol 584 (11) ◽  
pp. 2478-2484 ◽  
Author(s):  
Toshie Kambe ◽  
Tao Song ◽  
Tsuyoshi Takata ◽  
Naoya Hatano ◽  
Yoshiaki Miyamoto ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Active Site ◽  
Cysteine Residue ◽  
Dependent Protein Kinase ◽  
Active Site Cysteine ◽  
Dependent Protein ◽  
Active Site Cysteine Residue

scholarly journals Evidence for histidine in the active site of papain

1968 ◽  
Vol 108 (5) ◽  
pp. 855-859 ◽  
Author(s):  
S. S. Husain ◽  
G. Lowe
Keyword(s):  
Active Site ◽  
Tertiary Structure ◽  
Cysteine Residue ◽  
Optical Rotatory Dispersion ◽  
Performic Acid ◽  
Histidine Residue ◽  
Active Site Cysteine ◽  
Similar Amino Acid ◽  
Inhibited Enzyme ◽  
Active Site Cysteine Residue

Papain was irreversibly inhibited by 1,3-dibromoacetone, a reagent designed to react first with the active-site cysteine residue and subsequently with a second nucleophile. The molecular weight of the inhibited enzyme was indistinguishable from that of papain itself, and no evidence of dimeric or oligomeric species was found. The optical-rotatory-dispersion curves of chloroacetone-inhibited papain and 1,3-dibromoacetone-inhibited papain were essentially similar. Amino acid analysis of the 1,3-dibromo[2−14C]acetone-inhibited enzyme and the performic acid-oxidized material clearly showed that a cysteine and histidine residue had been alkylated through the thiol and N-1 of the imidazole group respectively. These groups must therefore be within 5å of each other in the tertiary structure of papain. Possible mechanistic implications are briefly discussed.

scholarly journals Distinct regulation of Ubc13 functions by the two ubiquitin-conjugating enzyme variants Mms2 and Uev1A

2005 ◽  
Vol 170 (5) ◽  
pp. 745-755 ◽  
Author(s):  
Parker L. Andersen ◽  
Honglin Zhou ◽  
Landon Pastushok ◽  
Trevor Moraes ◽  
Sean McKenna ◽  
...  
Keyword(s):  
Nuclear Factor Κb ◽  
Cysteine Residue ◽  
Physical Interaction ◽  
Polyubiquitin Chains ◽  
Active Site Cysteine ◽  
Cellular Processes ◽  
Damage Repair ◽  
Ubiquitin Conjugating Enzyme ◽  
Active Site Cysteine Residue ◽  
Conjugating Enzyme

Ubc13, a ubiquitin-conjugating enzyme (Ubc), requires the presence of a Ubc variant (Uev) for polyubiquitination. Uevs, although resembling Ubc in sequence and structure, lack the active site cysteine residue and are catalytically inactive. The yeast Uev (Mms2) incites noncanonical Lys63-linked polyubiquitination by Ubc13, whereas the increased diversity of Uevs in higher eukaryotes suggests an unexpected complication in ubiquitination. In this study, we demonstrate that divergent activities of mammalian Ubc13 rely on its pairing with either of two Uevs, Uev1A or Mms2. Structurally, we demonstrate that Mms2 and Uev1A differentially modulate the length of Ubc13-mediated Lys63-linked polyubiquitin chains. Functionally, we describe that Ubc13–Mms2 is required for DNA damage repair but not nuclear factor κB (NF-κB) activation, whereas Ubc13–Uev1A is involved in NF-κB activation but not DNA repair. Our finding suggests a novel regulatory mechanism in which different Uevs direct Ubcs to diverse cellular processes through physical interaction and alternative polyubiquitination.

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