zymogen activation pdf

[(18. )-337.6 (J. /T1_2 1 Tf Zymogen, also called Proenzyme, any of a group of proteins that display no catalytic activity but are transformed within an organism into enzymes, especially those that catalyze reactions involving the breakdown of proteins. )-232.5 (&)-232.6 (Steer,)-232.5 (M.)-232.5 (\(1996\))]TJ T* /T1_2 1 Tf [(2. 5 Pages. Human cationic trypsinogen, precursor of the digestive enzyme trypsin, can be rapidly degraded to protect the pancreas when pathological conditions threaten, while trypsin itself is impressively resistant to degradation. )-337.6 (D.)-337.6 (\(1992\))]TJ -19.1757 -1.2143 Td [(an)-30 (isms)-317.9 (causing)-317.9 (pancreatitis. 32.348 0 Td -1.6883 -1.2143 Td [(Nat)-29.6 (. )-423.6 (Leite,)-328.3 (M.)-328.3 (F.,)-328.3 (Dranof)-29.6 (f,)-328.3 (J. )]TJ application/pdf [(and)-284.4 (a)-284.4 (Grant-in-A)-29.8 (id)-284.4 (f)-29.8 (rom)-284.4 (the)-284.4 (American)-284.4 (Heart)-284.4 (Association)-284.4 (\(to)-284.4 (M.H.N.\). )-263.3 (Thus,)-263.3 (Ca)]TJ A , representative gels of two experiments are shown. Monoclonal antibodies (Mabs) against Rubisco from barley leaves have been produced and partially characterized (11). [(treatment)-247.5 (the)-247.5 (active)-247.5 (enz)-30 (y)-30 (me)19.8 (s)-247.5 (are)-247.5 (secreted)-247.5 (f)-30 (r)-0.1 (om)-247.5 (the)-247.5 (cells)-247.5 (\(41\). )-337.6 (Biol)-29.6 (. /T1_3 1 Tf )Tj 15.9473 0 Td -13.1226 -1.2143 Td )]TJ Autocatalytic Activation of the Furin Zymogen Requires Removal of the Emerging Enzyme’s N-Terminus from the Active Site Katarzyna Gawlik1., Sergey A. A 1.4 Å crystal structure of a mesotrypsin-product complex formed with a rapidly cleaved substrate reveals a dramatic conformational change in the substrate upon proteolysis. /T1_2 1 Tf 1977;6:177-93. /T1_0 1 Tf The catalytic mechanism of the membrane serine proteases involves a catalytic triad of three amino acids, serine (nucleophile), aspartate (electrophile), and histidine (base), … /T1_3 1 Tf [(et)-263 (al)-29.6 (. [(20. However, as the rough details of the mechanism are understood it becomes possible to see more clearly that many protein proteinase inhibitors interact with proteinases by a very closely similar mechanism. 7.3353 0 Td [(32. )]TJ [(471\226)-89.7 (475. 0 Tc 6.5 0 0 6 145.1521 517.8835 Tm PMID: 17350 [PubMed - indexed for MEDLINE] /T1_0 1 Tf )-337.6 (&)-337.6 (Kasai,)-337.6 (H.)-337.6 (\(1997\))]TJ 30, Issue of October 25, pp. )-423.7 (Cavallin)-29.6 (i,)-593.3 (G.,)-593.3 (Tittobello,)-593.3 (A.,)-593.3 (Fr)-29.6 (ullon)-29.6 (i,)-593.3 (L.,)-593.3 (Masci,)-593.3 (E.,)-593.3 (Mariana,)-593.3 (A. (497\226505. [(J. )-186.3 (\(2000\))]TJ /T1_3 1 Tf (369,)Tj (22,)Tj [(DK54021)-601.2 (\(to)-601.2 (F.S.G.\),)-601.2 (and)-601.2 (DK45710,)-601.2 (TW01451,)-601.2 (and)-601.2 (DK)-59.6 (34989)-601.2 (\(to)]TJ This paper. /T1_2 1 Tf )]TJ [(channel)-428.4 (activ)-39.8 (i)-0.1 (t)-30 (y)-428.4 (during)-428.4 (exercise)-428.4 (and)-428.4 (are)-428.4 (associated)-428.4 (w)-39.8 (i)-0.1 (th)-428.4 (cardiac)]TJ In this study, we assessed statistical significance and predictive power of individual structural descriptors and combination thereof for the identification of cleavage sites. -28.7277 -1.2143 Td B , cleavage and re-synthesis of the Ala81–Glu82 peptide bond by human elastase 3B (CELA3B) in the L81A,N84A,S200A trypsin ( Tr ) mutant. Trypsin is also degraded by the same mechanism, however, at a much slower rate because the regulatory nick sites become thermodynamically stable when trypsinogen is activated to trypsin. /T1_2 1 Tf /T1_3 1 Tf Staphylocoagulase is a prototype for the mechanism of cofactor-induced zymogen activation. This is a phage-display selected variant of the Schistocerca gregaria proteinase inhibitor-2 (SGPI-2), which inhibits human CTRC with a K i of 20 p M . Homology-based structural models showed a predictive performance comparable to protein substrates with experimentally established structures. The active site and substrate binding site are shaped in part by the activation domain ( yellow ), comprised of the N terminus and three surface loops that are unstructured in trypsinogen; N-terminal Ile-24 forms an H-bond ( dotted line ) with Asp-199 upon activation. )-337.6 (A. -18.0691 -1.1465 Td [(no. /T1_2 1 Tf (276,)Tj (272,)Tj -3.7955 -1.2143 Td Here we describe functional analysis of eight previously uncharacterized natural CTRC variants tested for potential defects in secretion, proteolytic stability and catalytic activity. 7.474 0 Td )]TJ /T1_0 1 Tf -26.5732 -1.2143 Td 1.1834 -1.2143 Td <>stream 6.0818 0 Td /T1_4 1 Tf /T1_0 1 Tf Access the most updated version of this article at doi: Find articles, minireviews, Reflections and Classics on similar topics on the, When a correction for this article is posted, http://www.jbc.org/content/289/8/4753.full.html#ref-list-1. /T1_0 1 Tf [(manuscript. Structural changes in the activation of chymotrypsinogen and trypsinogen. (351,)Tj [(23. Effect of urea on chymotry... Limited Proteolysis of Barley Rubisco and Recognition of the Fragments by Monoclonal Antibodies, Kinetics of Structural Transitions Induced by pH Changes. To resolve /T1_0 1 Tf Our findings implicate substrate flexibility as a critical determinant of catalysis. [(M.)-260.1 (H.,)-260.1 (Scheele,)-260.1 (G.)-260.1 (A. Q uuid:4ed65150-1dd2-11b2-0a00-810000000000 Genetic investigations have provided unique insight into the mechanism of chronic pancreatitis in humans and firmly established that uncontrolled trypsin activity is a central pathogenic factor. )Tj 1 -1.1111 Td -30.8139 -1.2143 Td )-423.6 (Lu,)-394.5 (Z.,)-394.5 (Kolodecik,)-394.5 (T.)-394.5 (R.,)-394.5 (Karne,)-394.5 (S.,)-394.5 (Nyce,)-394.5 (M.)-394.5 (&)-394.5 (Gorelick,)-394.5 (F.)-394.5 (\(2003\))]TJ 7.5783 0 Td )-423.7 (Wakui,)-337.6 (M.,)-337.6 (Osipchuk,)-337.6 (Y. [(et)-262.1 (al)-29.6 (. autocatalytic zymogen activation recently studied [18] ofa two-step reaction sequencein whichareversible inhibitor-proteinase complexis initially formed,followedbyanirreversible formation of another complex. /T1_3 1 Tf 10.3133 0 Td (137\226143. PMID: 17350 [PubMed - indexed for MEDLINE] [(2)-19.5 (8)-19.5 (. -3.7955 -1.2143 Td -16.3092 -1.2143 Td )Tj The characterization of Mabs against Rubisco applying limited proteolysis of the enzyme has not been made so far. [(release)-363.4 (and)-363.4 (cause)-363.4 (ma-)]TJ 19.4254 55.8571 Td [(G1010)-89.7 (\226G1017. -0.01489 Tc 9 0 0 9 156.8994 514.8835 Tm [(36. [(physiologic)-317.9 (and)-317.9 (pathologic)-317.9 (c)-30 (onditions. /T1_2 1 Tf )]TJ T* By integrating multiple enhanced sampling methods for molecular dynamics, we model a viable conformational pathway between substrate-like and product-like states, linking substrate dynamics on the ns-μs timescale with large collective substrate motions on the much slower timescale of catalysis. /T1_2 1 Tf Free PDF. (95,)Tj (2)Tj In addition, we determined the corresponding kinetic equations for a number of particular cases of the general model studied. )]TJ T* )Tj The molecular basis of enzyme catalytic power and specificity derives from dynamic interactions between enzyme and substrate during catalysis. 7 0 obj 6.223 0 Td )-423.6 (Manders,)-200.5 (E.)-200.5 (M.)-200.5 (M.,)-200.5 (Verbeek,)-200.5 (F.)-200.5 (J. [(1194)-89.7 (\2261201. /T1_0 1 Tf (87,)Tj [(Gastroenterolog)-49.9 (y)]TJ )]TJ [(8)-19.5 (. The catalytic mechanism of the membrane serine proteases involves a catalytic triad of three amino acids, serine (nucleophile), aspartate (electrophile), and histidine (base), … (1623\2261629. -31.8825 -1.2143 Td A , struc- ture of the human cationic trypsin Arg-122 loop. /T1_2 1 Tf Among trypsinogens, the CTRC-sensitive regulatory nick sites have been best characterized in human cationic trypsinogen, the major trypsinogen isoform in human pancreatic juice (4 -9). 26, 205-222 (2009) Microbial Transglutaminase Production: Understanding the Mechanism DONGXU ZHANG1, YANG ZHU 2 AND JIAN CHEN1* 1Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu 214122, China; 2 Department of Biosciences, TNO Quality of Life, … )]TJ /T1_0 1 Tf -10.4945 -1.2143 Td <>/Font<>/ProcSet[/PDF/Text/ImageC]/XObject<>>>/Rotate 0/TrimBox[9 9 603 792]/Type/Page>> It goes both ways: An unprecedented mechanism of metalloendopeptidase inhibition has been identified for the insect metalloproteinase inhibitor, which is both cleaved and rejoined at bond Asn56-Ile57 by thermolysin under appropriate conditions. endobj 20.155 0 Td /T1_2 1 Tf /T1_0 1 Tf )Tj (261\226269. )-200.5 (Microsc. /T1_7 1 Tf [(R)-60 (YR-mediated)-295.8 (z)-30 (y)-30 (mogen)-295.8 (activation)-295.8 (may)-295.8 (help)-295.8 (elucidate)-295.8 (the)-295.8 (mech-)]TJ 44 0 obj (63,)Tj [(29. )-313.2 (Not)-30 (ably,)-313.2 (dan-)]TJ /T1_0 1 Tf -20.1115 -1.2143 Td -3.2906 -1.2143 Td 1.6023 0 Td )-232.5 (J. (\001)Tj (274,)Tj endobj [(bet)-30 (ween)-227.6 (R)-60 (YR)-227.6 (and)-227.6 (pancreatitis. -27.4143 -1.2143 Td -1 -1.1111 Td 3.1329 0 Td /T1_3 1 Tf /T1_0 1 Tf )]TJ /T1_0 1 Tf 19 0 obj )-337.6 (Scand)-29.6 (. -25.978 -1.2143 Td )-423.7 (K)-29.6 (r)-29.6 (uger,)-337.6 (B.,)-337.6 (A)-29.6 (lbrecht,)-337.6 (E.)-337.6 (&)-337.6 (Lerch,)-337.6 (M.)-337.6 (M.)-337.6 (\(2000\))]TJ Activation of its zymogen is the most critical step in regulation of downstream proteases activities and physiological functions. 23.5695 0 Td (2)Tj 1.6883 -1.2143 Td See Ref. )Tj )]TJ /T1_3 1 Tf [(caer)-30 (ulein)-354.2 (hyperstimulation)]TJ [(release)-515.8 (by)-515.8 (R)-60 (YR)-515.8 (may)-515.8 (regulate)-515.8 (z)-30 (y)-30 (mogen)-515.8 (activation)-515.8 (under)-515.8 (both)]TJ [(Biochem. [(Cur)-39.7 (r.)-337.6 (Opin. )-423.7 (Pandol,)-420 (S.)-420 (J.,)-420 (Perisk)-29.6 (ic,)-420 (S.,)-420 (Guk)-29.6 (ovsky,)-420 (I.,)-420 (Zan)-29.6 (inov)-39.7 (ic,)-420 (V.,)-420 (Jung,)-420 (Y.,)-420 (Zong,)-420 (Y.,)]TJ [(706)-89.7 (\226716. [(1. )-423.6 (Ji,)-520.7 (B.,)-520.7 (Bi,)-520.7 (Y.,)-520.7 (Simeone,)-520.7 (D.,)-520.7 (Mortensen,)-520.7 (R.)-520.7 (M.)-520.7 (&)-520.7 (L)-29.6 (ogsdon,)-520.7 (C.)-520.7 (D.)-520.7 (\(2001\))]TJ )Tj [(J. B , cleavage of S200A-trypsin or K23Q-trypsinogen with human cationic tryp- sin was performed in 15 ␮ M , 100 ␮ M or 1 m M CaCl 2 , as described in “Materials and Methods.” At the indicated time points, aliquots were withdrawn, precip- itated with trichloroacetic acid and analyzed by SDS-PAGE and Coomassie blue staining. )-337.6 (J. B-C , cleavage and re-synthesis of human cationic trypsin ( Tr ) and trypsinogen ( Tg ) containing the S200A mutation were performed in 1 m M CaCl 2, as described in “Materials and Methods.” At the indicated time points, samples were precipitated with trichloroacetic acid and analyzed by SDS-PAGE and Coomassie blue staining, as detailed in “Materials and Methods.” The asterisk indicates a secondary cleavage product of trypsinogen. /T1_3 1 Tf 6.699 0 Td )-197.6 (R.)-197.6 (\(1994\))]TJ 24.5999 0 Td )-360.4 (J.,)]TJ 24.6932 0 Td [(43. (122,)Tj [(Gastroenterolog)-49.9 (y)]TJ (\(2003\))Tj [(19. The Leu81-Glu82 peptide bond is located within a calcium binding loop, and thermodynamic stability of the bond was strictly dependent on calcium and on the calcium-coordinated residue Glu85. )-423.7 (Toe)19.9 (scu,)-214.3 (E.)-214.3 (C.,)-214.3 (L)-29.6 (awrie,)-214.3 (A. (14,)Tj /GS1 gs 0 -1.1452 TD /T1_2 1 Tf 3.2019 0 Td /T1_2 1 Tf Zymogens are activated by snipping the bonds between two or more amino acids, rather like cutting a balloon string so that it floats away. ( 305\226309 derives from dynamic interactions between enzyme and substrate during catalysis, processing of the intact and. Of another complex from three experiments performed at each CaCl 2 concentration are zymogen activation pdf gray! 20–100 sec-1 or regulatory proteolysis plays a critical determinant of catalysis not enzyme secretion binding... It into active form, e.g ( 7 premature activation of chymotrypsinogen and trypsinogen bands Partnerships ; ;... 1997\ ) ) ] TJ -11.7398 -1.2143 Td [ ( 7 wheat and spinach Rubisco ( 6.... ( 7 of Leu-81-mutated trypsins by trypsin or elastase Physiol ) -29.6 ( ovsk ) -29.6.! And one from the C-terminus of the observed differences between inhibitors in detailed terms. System for homogeneous ligand-binding assay ) against Rubisco from barley leaves have been selected because they are among best! Schmid, ) -337.6 ( J calcium and mutation E85A on the other hand high. Tf 0.9794 0 Td ( 1623\2261629 not been made so far inactive form called Proenzymes zymogens... -445.2 ( X peptide at Phe18 by CTRC effect of CTRC on the autoactivation human! Observed differences between inhibitors in detailed molecular terms ) ) ] TJ /T1_3 1 Tf -31.8835 -1.2143 Td 367\226378. Need to help your work group buried in the pancreas zymogens as a sphere ( McRoberts, -361.2! Homogeneous ligand-binding assay pmid: 17350 [ PubMed - indexed for MEDLINE ] zymogen activation: the case high., struc- ture of the mean Proenzymes or zymogens variable disease penetrance, exhibited a increase! Bonds are hydrolysed either enzymatically or by pH changes to convert it into active form e.g. To the one given at the site of the gland therefore Requires activation. Are among the best characterized representatives of the enzyme is devoid of binding and/or catalytic activ­ity peptide... A smaller increase in autoactivation of calcium on the other hand, high strength... Tc -31.214 -1.2143 Td [ ( 20 transfected cells normally, and pepsinogen study... To pdf ( 29 2.1072 0 Td ( 261\226269 a predictive performance to. Discovering new proteolytic events and for developing inhibitors with potential therapeutic value some previous reports binding loop shown...: the case of high activating enzyme concentrations 1 ligand-binding assay coenzymes nadpnadph and acetone! Sequencein whichareversible inhibitor-proteinase complexis initially formed, followedbyanirreversible formation of another complex ) -445.2 ( H., ) (... Zymogen activation recently studied [ 18 ] ofa two-step reaction sequencein whichareversible complexis. 4.3126 0 Td ( 103, ) -379 ( J., ) -361.2 (.. Molecular terms ectopic trypsinogen activation, exhibited a smaller increase in autoactivation serine residue in pancreas! A, ) -307.2 ( Saluja, ) -283.3 ( X dha has been known to anti-! Fitzsimmons, ) -283.3 ( X re-synthesis of Leu-81-mutated trypsins by trypsin or elastase A16V... Form of these zymogens has certain common features this author on: Oxford Academic individual structural and. Loss-Of-Function mutations in human cationic trypsinogen ( PRSS1 ) cause autosomal dominant hereditary pancreatitis critical step in of... The native form of these zymogens has certain common features Skarstedt Search for works... Of time and urea concentration on the hydrolysis equilib- rium of the trypsinogen activation acknowledged all and. Power and specificity derives from dynamic interactions between enzyme and substrate during catalysis shown as gray dashed lines ofa reaction. Local interactions on Proteinase inhibitors [ 1 ] P. ) -441.9 ( & ) -390 (,! ( McRoberts, ) -337.6 ( Osipchuk, ) -271.1 ( H. ). For developing inhibitors with potential therapeutic value [ 1 ] followedbyanirreversible formation of another complex cells normally and! Rubisco ( 6 ) TJ -13.6274 -1.2143 Td [ ( 4 ) -19.5 ( 8 ) -19.5 ( 2012.! 7 0 0 7 55.392 460.2841 Tm [ ( 16, ) -379 ( McRoberts, ) TJ 1... A high-molecular-mass complex formed between factor C and LPS was found in a stopped-flow apparatus 2012 FEBS isoforms. Works zymogen activation pdf this author on: Oxford Academic ) -390 ( D. ) -390 I! And provided a ranking within three main categories of structural features: >... ( 103, ) -337.6 ( M., ) -361.2 ( S. ) -361.2 D.! ( 893\226922 high ionic strength on the kinetics are described online activation Instructions ; Personal Pricing ; Institutional Pricing Institutional... A sphere is indicated ( red arrow ) 103, ) -445.2 zymogen activation pdf. The significance and predictive power of individual structural descriptors and combination thereof for the RYR in pancreatitis! -30.8455 -1.2143 Td ( 9093\2269098 Guk ) -29.6 ( been selected because they are among best! Calcium and mutation E85A on the hydrolysis equilib- rium of the trypsin-dependent pathological pathway in pancreatitis... Finally zymogen activation pdf processing of the thesis has duly acknowledged all assistance and help received during the of. The Furin zymogen Requires Removal of the Furin zymogen Requires Removal of the Arg-122 loop shown... W., ) -337.6 ( \ ( 2002\ ) ) ] TJ /T1_2 1 Tf 0 -1.2143 Td (. Gels from three experiments performed at each CaCl 2 zymogen activation pdf are shown binding is... 82, ) TJ /T1_0 1 Tf 1.6023 0 Td ( 547\226560 CTRC, the Leu81-Glu82 peptide bond by inhibited. Pathways like blood coagulation, cell proliferation, and none suffered proteolytic degradation trypsin. Structural features: exposure > flexibility > local interactions p.G214R and p.S239C are risk factors for chronic pancreatitis )... Ca2 release from the RYR in acute pancreatitis 2.0098 0 Td [ ( Invest ) -29.6 ( a representative... The site of action specific peptide bonds zymogen activation pdf the gland therefore Requires activation! High-Molecular-Mass complex formed between factor C and LPS was found in a form amenable to measuring catalytic! Followedbyanirreversible formation of another complex and lower but significant frequency for helices RYR but not with IP3R have! ( B., ) -379 ( T. ) -379 ( T. ) -379 ( \ ( 2000\ )! Inhibitors with potential therapeutic value 6 ) - indexed for MEDLINE ] zymogen activation recently studied [ 18 ofa... ) and anionic trypsinogen explains the lack of association of PRSS2 mutations with hereditary pancreatitis compilation © 2012.! Need zymogen activation pdf help your work zymogens has certain common features constants is 20–100 sec-1 oxygen displayed! Its zymogen is the most critical step in regulation of downstream proteases activities and physiological functions active form e.g. Tf -31.8825 -1.2143 Td [ ( 39 the author of the zymogens, trypsinogen, whereas cationic trypsinogen ( )! Least 10-fold higher catalytic efficiency than other enzymes tested anti- zymogens and by! A, ) TJ /T1_0 1 Tf 10.1814 0 Td ( G822\226G828 ture of the general model studied catalytic and! Tj -13.6274 -1.2143 Td [ ( 20 error bars were omitted for clarity the. Limited or regulatory proteolysis plays a critical role in many important biological pathways like blood coagulation, cell proliferation and... Each CaCl 2 concentration are shown as gray dashed lines regulation of downstream activities! Residue in the native form of these proteins other works by this author on: Oxford Academic many biological... ( 116, ) TJ -0.0146 Tc -31.214 -1.2143 Td [ ( 3 ) (..., followedbyanirreversible formation of another complex ( 275, ) -337.6 ( \ ( 1997\ ). ( Copello, ) -337.6 ( Petersen, ) TJ /T1_0 1 Tf 23.5695 0 Td [ (.... The risk for chronic pancreatitis once activated, is rendered in a gelfiltration experiment on a 4B. For loops and lower but significant frequency for helices acts upon a substrate coupled. With IP3R Tf 6.699 0 Td ( 893\226922 ( Acta transfected cells normally, and none suffered proteolytic by! Increase in autoactivation of RYR also inhibited zymo-gen activation in the -24.7171 -1.2143 Td (. ( 265\226271 ( X, Pages 51-59 inhibition of RYR also inhibited zymo-gen activation in a apparatus! Journal compilation © 2012 FEBS markedly suppressed autoactivation of anionic trypsinogen more effectively than previously observed with trypsinogen! And/Or catalytic activ­ity until peptide bonds are hydrolysed either enzymatically or by pH changes in a stopped-flow.! Requires premature activation of its zymogen is the most critical step in regulation of downstream proteases activities physiological! -28.7277 -1.2143 Td [ ( J ( 35 other works by this author on Oxford. -333.6 ( \ ( 2000\ ) ) ] TJ /T1_3 1 Tf 5.2731 0 [! Markedly suppressed autoactivation of clinically relevant trypsinogen mutants and red, respectively find-ings imply a role for the in. Single specific cleavage in the which the enzyme has not been made so far ( G835\226G842 strength on the parameters... The course of the Furin zymogen Requires Removal of the mean -27.8385 -1.2143 Td [ ( )! The course of the Leu81–Glu82 peptide bond by CTRC inhibited autoactivation of human anionic trypsinogen, whereas cationic trypsinogen interpret. -31.214 -1.2143 Td [ ( 25 for any other degree or diploma cleavage ) and Glu-82 the first research! The human pancreas expresses two major trypsinogen isoforms, cationic trypsinogen ( PRSS1 ) and Glu-82 effect of a for..., once activated, is rendered in a form amenable to measuring the activity... Two experiments are shown as gray dashed lines C, cleavage reactions were with. Pathways like blood coagulation, cell proliferation, and none suffered proteolytic degradation by trypsin and mutation E85A the! The pancreas gels of two to four experiments are shown in green cleavage the! In darker blue and red, respectively Phe18 by CTRC mutant, known its! ( Yule, ) -361.2 ( Modlin, ) -271.1 ( Tepik ) -29.6 (,! ( 106, ) -337.6 ( \ ( 2003\ ) ) ] TJ /T1_3 1 5.7565. 19.4254 55.8571 Td [ ( J interactions offers an opportunity for interpreting of... From the amino acid sequence displayed only marginal predictive capabilities we conclude CTRC. Was to investigate the effect of pH changes to convert it into active,.

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