scholarly journals Evidence for histidine in the active sites of ficin and stem-bromelain

1968 ◽  
Vol 110 (1) ◽  
pp. 53-57 ◽  
Author(s):  
S. S. Husain ◽  
G. Lowe
Keyword(s):  
Active Site ◽  
Active Sites ◽  
Cysteine Residue ◽  
Histidine Residue ◽  
Active Site Cysteine ◽  
Stem Bromelain ◽  
Hydrolysis Of ◽  
Active Site Cysteine Residue

1. Ficin and stem-bromelain are irreversibly inhibited by 1,3-dibromoacetone, a reagent designed to react first with the active-site cysteine residue and subsequently with a second nucleophile. Evidence is presented that establishes that a histidine residue is within a 5Å locus of the active-site cysteine residue in both enzymes. The histidine residue in both enzymes is alkylated at N-1 by dibromoacetone. It is suggested that, as with papain, the thiol and imidazole groups act in concert in the hydrolysis of substrates by these enzymes. 2. The inhibition of thiol-subtilisin with 1,3-dibromoacetone is shown to be due to the alkylation of a cysteine residue only.

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 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 The electrostatic fields in the active-site clefts of actinidin and papain

1988 ◽  
Vol 254 (1) ◽  
pp. 235-238 ◽  
Author(s):  
R W Pickersgill ◽  
P W Goodenough ◽  
I G Sumner ◽  
M E Collins
Keyword(s):  
Active Site ◽  
Active Sites ◽  
Structural Similarity ◽  
Cysteine Residue ◽  
Site Directed Mutagenesis ◽  
Activity Profile ◽  
Active Site Cysteine ◽  
Electrostatic Fields ◽  
Imidazolium Ion ◽  
Active Site Cysteine Residue

The active sites of actinidin (EC 3.4.22.14) and papain (EC 3.4.22.2) display different reactivity characteristics to probes targeted at the active-site cysteine residue despite the close structural similarity of their active sites. The calculated electrostatic fields in the active-site clefts of actinidin and papain differ significantly and may explain the reactivity characteristics of these enzymes. Calculation of electrostatic potential also focuses attention on the electrostatic properties that govern formation of the active-site thiolate-imidazolium ion-pair. These calculations will guide the modification of the pH-activity profile of the cysteine proteinases by site-directed mutagenesis.

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

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 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

Allosteric Inhibition of PTP1B Activity by Selective Modification of a Non-Active Site Cysteine Residue†

Biochemistry ◽  
2005 ◽  
Vol 44 (21) ◽  
pp. 7704-7712 ◽  
Author(s):  
Stig K. Hansen ◽  
Mark T. Cancilla ◽  
Timothy P. Shiau ◽  
Jenny Kung ◽  
Teresa Chen ◽  
...  
Keyword(s):  
Active Site ◽  
Cysteine Residue ◽  
Allosteric Inhibition ◽  
Active Site Cysteine ◽  
Active Site Cysteine Residue
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