00554nas a2200145 4500008004100000245009000041210006900131260000800200300001000208490000600218100002300224700002100247700002500268856011500293 2008 eng d00aIdentification of ripening specific proteins in tomato by intact tissue MALDI-TOF-MS.0 aIdentification of ripening specific proteins in tomato by intact cJAN a10-130 v31 aKorwar, Arvind, M.1 aSanthakumari, B.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/identification-ripening-specific-proteins-tomato-intact-tissue-maldi-tof-ms01657nas a2200265 4500008004100000022001400041245010500055210006900160260009500229300001200324490000800336520066400344653003601008653001801044653000901062100002301071700002501094700002401119700002101143700002201164700002201186700003301208700002501241856012501266 2012 eng d a0006-291X00aAnalysis of AGE modified proteins and RAGE expression in HER2/neu negative invasive ductal carcinoma0 aAnalysis of AGE modified proteins and RAGE expression in HER2neu a525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USAbACADEMIC PRESS INC ELSEVIER SCIENCEcMAR a490-4940 v4193 a

Cancer is associated with increased glycolysis and carbonyl stress. In view of this, AGE modified proteins were identified from clinical breast cancer tissue using 2DE-immunoblot and mass-spectrometry. These proteins were identified to be serotransferrin, fibrinogen gamma chain, glycerol-3-phosphate dehydrogenase, lactate dehydrogenase, annexin II, prohibitin and peroxiredoxin 6, which have established role in cancer. Further, RAGE expression and its downstream signaling proteins NADPH oxidase and NF-kB were studied. Role of these AGE modified proteins and RAGE signaling in breast cancer is discussed. (C) 2012 Elsevier Inc. All rights reserved.

10aadvanced glycation end products10aBreast cancer10aRAGE1 aKorwar, Arvind, M.1 aBhonsle, Hemangi, S.1 aChougale, Ashok, D.1 aKote, Sachin, S.1 aGawai, Kachru, R.1 aGhole, Vikram, S.1 aKoppikar, Chaitanyananda, B.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/analysis-age-modified-proteins-and-rage-expression-her2neu-negative-invasive-ductal-002403nas a2200313 4500008004100000022001400041245010700055210006900162260007100231300001400302490000700316520127100323653004301594653001201637653001201649653003601661100002501697700002301722700002101745700002901766700002401795700002301819700002201842700002001864700002701884700002601911700002501937856012701962 2012 eng d a1535-389300aLow plasma albumin levels are associated with increased plasma protein glycation and HbA1c in diabetes0 aLow plasma albumin levels are associated with increased plasma p a1155 16TH ST, NW, WASHINGTON, DC 20036 USAbAMER CHEMICAL SOCcFEB a1391-13960 v113 a

Albumin is one of the most abundant plasma proteins and is heavily glycated in diabetes. In this study, we have addressed whether variation in the albumin levels influence glycation of plasma proteins and HbA1c. The study was performed in three systems: (1) streptozotocin (STZ)-induced diabetic mice plasma, (2) diabetic clinical plasma, and (3) in vitro glycated plasma. Diabetic mice and clinical plasma samples were categorized as diabetic high albumin plasma (DHAP) and diabetic low albumin plasma (DLAP) on the basis of their albumin levels. For the in vitro experiment, two albumin levels, high albumin plasma (HAP) and low albumin plasma (LAP), were created by differential depletion of plasma albumin. Protein glycation was studied by using a combination of two-dimensional electrophoresis (2DE), Western blotting, and LC-MSE. In both mice and clinical experiments, an increased plasma protein glycation was observed in DLAP than in DHAP. Additionally, plasma albumin levels were negatively correlated with HbA1c. The in vitro experiment with differential depletion of albumin mechanistically showed that the low albumin levels are associated with increased plasma protein glycation and that albumin competes for glycation with other plasma proteins.

10aadvanced glycation end products (AGEs)10aAlbumin10aglucose10apost translational modification1 aBhonsle, Hemangi, S.1 aKorwar, Arvind, M.1 aKote, Sachin, S.1 aGolegaonkar, Sandeep, B.1 aChougale, Ashok, D.1 aShaik, Mahemud, L.1 aDhande, Nitin, L.1 aGiri, Ashok, P.1 aShelgikar, Kishore, M.1 aBoppana, Ramanamurthy1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/low-plasma-albumin-levels-are-associated-increased-plasma-protein-glycation-and-hba1c-002064nas a2200253 4500008004100000022001400041245013500055210006900190260009300259300001200352490000700364520103800371653004301409653003301452653001301485653003701498100002501535700002301560700002401583700002501607700002301632700002501655856013001680 2012 eng d a1469-066700aZoom-in - a targeted database search for identification of glycation modifications analyzed by untargeted tandem mass spectrometry0 aZoomin a targeted database search for identification of glycatio a6 CHARLTON MILL, CHARLTON, CHICHESTER,, W SUSSEX PO18 0HY, ENGLANDbIM PUBLICATIONScJAN a475-4810 v183 a

Post-translational modifications (PTMs) are very important to biological function, however their identification and characterization is technically challenging. In this study, we have identified glycation modifications by nano LC-MSE, a data independent acquisition work flow, followed by database search using the Protein Lynx Global Server (PLGS). PLGS search with a complete human protein database hardly identified glycation modifications in a glycated human serum albumin (HSA), which was detected to be glycated by western blotting with advanced glycation end products (AGE) antibody and fluorescence spectroscopy. To overcome this difficulty, ``Zoom-In'' approach, a targeted database search was used to identify glycation modifications in a glycated HSA, which were further manually validated. This approach was useful for identification of glycation modifications from untargeted tandem mass spectrometry workflow such as MSE, but may require the development of a new algorithm or an upgrade of the existing software.

10aadvanced glycation end products (AGEs)10adata independent acquisition10aDiabetes10apost-translational modifications1 aBhonsle, Hemangi, S.1 aKorwar, Arvind, M.1 aKesavan, Suresh, K.1 aBhosale, Santosh, D.1 aBansode, Sneha, B.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/zoom-targeted-database-search-identification-glycation-modifications-analyzed-untargeted-001301nas a2200241 4500008004100000022001400041245005300055210005200108260007500160300001200235490000600247520057000253653001000823653001300833653001400846653000900860100002500869700002300894700001600917700002500933700002000958856008100978 2013 eng d a1862-834600aGlycated proteome: from reaction to intervention0 aGlycated proteome from reaction to intervention aPOSTFACH 101161, 69451 WEINHEIM, GERMANYbWILEY-V C H VERLAG GMBHcJAN a155-1700 v73 a

Glycation, a nonenzymatic reaction between reducing sugars and proteins, is a proteome wide phenomenon, predominantly observed in diabetes due to hyperglycemia. Glycated proteome of plasma, kidney, lens, and brain are implicated in the pathogenesis of various diseases, including diabetic complications, neurodegenerative diseases, cancer, and aging. This review discusses the strategies to characterize protein glycation, its functional implications in different diseases, and intervention strategies to protect the deleterious effects of protein glycation.

10aaging10aDiabetes10aGlycation10aRAGE1 aKulkarni, Mahesh, J.1 aKorwar, Arvind, M.1 aMary, Sheon1 aBhonsle, Hemangi, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/glycated-proteome-reaction-intervention-002352nas a2200205 4500008004100000022001400041245015300055210006900208260008800277300001000365490000700375520148400382100002301866700002501889700002201914700002201936700003301958700002501991856013002016 2013 eng d a1536-231000aProteomic profiling and interactome analysis of ER-positive/HER2/neu negative invasive ductal carcinoma of the breast: towards proteomics biomarkers0 aProteomic profiling and interactome analysis of ERpositiveHER2ne a140 HUGUENOT STREET, 3RD FL, NEW ROCHELLE, NY 10801 USAbMARY ANN LIEBERT, INCcJAN a27-400 v173 a

Breast cancer, especially ER positive/HER2/neu negative IDC, is the predominant subtype of invasive ductal carcinoma. Although proteomic approaches have been used towards biomarker discovery in clinical breast cancer, ER positive/HER2/neu negative IDC is the least studied subtype. To discover biomarkers, as well as to understand the molecular events associated with disease progression of estrogen receptor positive/HER2/neu negative subtype of invasive ductal carcinoma, differential protein expression profiling was performed by using LC-MSE (MS at elevated energy). A total of 118 proteins were identified, of which 26 were differentially expressed. These identified proteins were functionally classified and their interactions and coexpression were analyzed by using bioinformatic tools PANTHER (Protein Analysis THrough Evolutionary Relationships) and STRING (Search Tool for the Retrieval of Interacting Genes). These proteins were found to be upregulated and were involved in cytoskeletal organization, calcium binding, and stress response. Interactions of annexin A5, actin, S100 A10, glyceraldehyde 3 phosphate dehydrogenase, superoxide dismutase 1, apolipoprotein, fibrinogen, and heat shock proteins were prominent. Differential expression of these proteins was validated by two-dimensional gel electrophoresis and Western blot analysis. The cluster of these proteins may serve as a signature profile for estrogen receptor positive/ HER2/neu negative subtype.

1 aKorwar, Arvind, M.1 aBhonsle, Hemangi, S.1 aGhole, Vikram, S.1 aGawai, Kachru, R.1 aKoppikar, Chaitanyananda, B.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/proteomic-profiling-and-interactome-analysis-er-positiveher2neu-negative-invasive-ductal-001879nas a2200253 4500008004100000022001400041245011900055210006900174260006300243300001200306490000600318520096300324653001401287653001001301653001501311100002501326700002301351700002401374700002101398700002201419700002701441700002501468856013201493 2013 eng d a1791-299700aProteomic study reveals downregulation of apolipoprotein A1 in plasma of poorly controlled diabetes: a pilot study0 aProteomic study reveals downregulation of apolipoprotein A1 in p aPOB 18179, ATHENS, 116 10, GREECEbSPANDIDOS PUBL LTDcFEB a495-4980 v73 a

Proteomic approaches aid in gaining a better understanding of the pathophysiology of diabetic complications. In view of this, differential protein expression in diabetic plasma samples was studied by a combination of proteomic and western blot analyses. Diabetic plasma samples were categorized based on glycated haemoglobin levels as controlled diabetes (CD; 7-8%), poorly controlled diabetes (PCD; >8%) and non-diabetic control (ND;<6.4%). Two-dimensional electrophoresis and liquid chromatography-mass spectrometry revealed differential expression of proteins including upregulation of fibrinogen and haptoglobin and downregulation of vitamin D binding protein, alpha-1-antitrypsin, transthyretin and apolipoprotein A1 (Apo A1) in diabetic compared with non-diabetic plasma samples. Amongst these proteins, Apo A1 downregulation was prominent in PCD. Downregulation of Apo A1 may serve as an early predictive marker of diabetic complications.

10abiomarker10aHbA1c10aproteomics1 aBhonsle, Hemangi, S.1 aKorwar, Arvind, M.1 aChougale, Ashok, D.1 aKote, Sachin, S.1 aDhande, Nitin, L.1 aShelgikar, Kishore, M.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/proteomic-study-reveals-downregulation-apolipoprotein-a1-plasma-poorly-controlled-diabetes-002633nas a2200265 4500008004100000022001400041245022300055210006900278260010300347300001400450490000700464520147500471100002301946700002901969700003901998700002902037700001702066700002502083700002702108700002002135700002302155700003202178700002502210856013202235 2015 eng d a1535-947600aDevelopment of diagnostic fragment ion library for glycated peptides of human serum albumin: targeted quantification in prediabetic, diabetic, and microalbuminuria plasma by parallel reaction monitoring, SWATH, and MSE0 aDevelopment of diagnostic fragment ion library for glycated pept a9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USAbAMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INCcAUG a2150-21590 v143 a

Human serum albumin is one of the most abundant plasma proteins that readily undergoes glycation, thus glycated albumin has been suggested as an additional marker for monitoring glycemic status. Hitherto, only Amadori-modified peptides of albumin were quantified. In this study, we report the construction of fragment ion library for Amadori-modified lysine (AML), N(epsilon)-(carboxymethyl) lysine (CML)-, and N(epsilon)-(carboxyethyl) lysine (CEL)-modified peptides of the corresponding synthetically modified albumin using high resolution accurate mass spectrometry (HR/AM). The glycated peptides were manually inspected and validated for their modification. Further, the fragment ion library was used for quantification of glycated peptides of albumin in the context of diabetes. Targeted Sequential Window Acquisition of all THeoretical Mass Spectra (SWATH) analysis in pooled plasma samples of control, prediabetes, diabetes, and microalbuminuria, has led to identification and quantification of 13 glycated peptides comprised of four AML, seven CML, and two CEL modifications, representing nine lysine sites of albumin. Five lysine sites namely K549, K438, K490, K88, and K375, were observed to be highly sensitive for glycation modification as their respective m/z showed maximum fold change and had both AML and CML modifications. Thus, peptides involving these lysine sites could be potential novel markers to assess the degree of glycation in diabetes.

1 aKorwar, Arvind, M.1 aVannuruswamy, Garikapati1 aJagadeeshaprasad, Mashanipalya, G.1 aJayaramaiah, Ramesha, H.1 aBhat, Shweta1 aRegin, Bhaskaran, S.1 aRamaswamy, Sureshkumar1 aGiri, Ashok, P.1 aMohan, Viswanathan1 aBalasubramanyam, Muthuswamy1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/development-diagnostic-fragment-ion-library-glycated-peptides-human-serum-albumin-targeted-002083nas a2200181 4500008004100000022001400041245008000055210006900135260011000204300001600314490000600330520136100336100002901697700002301726700002501749700002101774856010601795 2015 eng d a2046-206900aRecent development of plant products with anti-glycation activity: a review0 aRecent development of plant products with antiglycation activity aTHOMAS GRAHAM HOUSE, SCIENCE PARK, MILTON RD, CAMBRIDGE CB4 0WF, CAMBS, ENGLANDbROYAL SOC CHEMISTRYcMAR a31113-311380 v53 a

Diabetes mellitus (DM) is an endocrine disorder characterized by chronic hyperglycemia, which results from an absolute or a relative deficiency of insulin or resistance to insulin. Hyperglycemia is increasingly linked to the pathogenesis of diabetic complications in individuals with long-duration diabetes. One of the inevitable consequences of hyperglycemia is the enhanced accumulation of advanced glycation end-products (AGEs), which are implicated in the pathogenesis of diabetes. Various natural products and their active constituents have reportedly been used for the treatment of diabetes and its complications. Some of these molecules are known to have anti-glycation activity. The search for novel anti-glycation agents from various sources is gaining a lot of importance. Attention has especially been focused on plants with an ethnopharmacological background and also on plants rich in triterpenoids and phenolics, which generally exhibit antioxidant and anti-glycation effects. Plant extracts or compounds obtained from them that possess both antioxidant and anti-glycation activities might have great therapeutic potential for treating diabetic complications. This review highlights the anti-glycation activities of phytochemicals, which will aid in the identification of lead molecules for the development of new anti-glycation drugs.

1 aChinchansure, Ashish, A.1 aKorwar, Arvind, M.1 aKulkarni, Mahesh, J.1 aJoshi, Swati, P. uhttps://library.ncl.res.in/content/recent-development-plant-products-anti-glycation-activity-review-002204nas a2200253 4500008004100000022001400041245009100055210006900146260011000215300001600325490000600341520123900347100002901586700003901615700001801654700002401672700001701696700002301713700002401736700002601760700002101786700002501807856011801832 2016 eng d a2046-206900aMolecules with O-acetyl group protect protein glycation by acetylating lysine residues0 aMolecules with Oacetyl group protect protein glycation by acetyl aTHOMAS GRAHAM HOUSE, SCIENCE PARK, MILTON RD, CAMBRIDGE CB4 0WF, CAMBS, ENGLANDbROYAL SOC CHEMISTRYcJUL a65572-655780 v63 a

Pharmaceutical intervention for reduction of advanced glycation end products (AGEs) is considered as a therapeutic strategy to attenuate the pathogenesis of diabetes. Many molecules have been reported to possess antiglycation activity, one such example is acetylsalicylic acid (aspirin). It protects proteins from glycation by acetylating the lysine residues. Therefore, in this study we have synthesized and screened molecules containing free N-acetyl, O-acetyl and acetophenone groups. All the selected molecules in this study showed glycation inhibition but interestingly, only molecules with O-acetyl but not N-acetyl and acetophenone groups were capable of acetylating lysine residue. Furthermore, we have demonstrated that pre-acetylation or aspirin treatment prior to the induction of diabetes helps in reducing HbA1c and AGE formation in the streptozotocin induced diabetic mice. Hence pre-acetylation may have an additional therapeutic efficacy of reducing AGE levels in vivo. Incorporation of O-acetyl group into anti-diabetic molecules could be a useful strategy, as it may have an additive effect in reducing AGEs. Identification of such novel acetylating agents represents a new area in the drug discovery process.

1 aVannuruswamy, Garikapati1 aJagadeeshaprasad, Mashanipalya, G.1 aKashinath, K.1 aKesavan, Suresh, K.1 aBhat, Shweta1 aKorwar, Arvind, M.1 aChougale, Ashok, D.1 aBoppana, Ramanamurthy1 aReddy, Srinivasa1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/molecules-o-acetyl-group-protect-protein-glycation-acetylating-lysine-residues02556nas a2200277 4500008004100000022001400041245012900055210006900184260009000253490000600343520147700349100002001826700002901846700002401875700002901899700002301928700001601951700002601967700002401993700002202017700002602039700002502065700003702090700002002127856013102147 2016 eng d a2045-232200aPotential dual role of eugenol in inhibiting advanced glycation end products in diabetes: proteomic and mechanistic insights0 aPotential dual role of eugenol in inhibiting advanced glycation  aMACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLANDbNATURE PUBLISHING GROUPcJAN0 v63 a

Medicinally important genus Ocimum harbors a vast pool of chemically diverse metabolites. Current study aims at identifying anti-diabetic candidate compounds from Ocimum species. Major metabolites in O. kilimandscharicum, O. tenuiflorum, O. gratissimum were purified, characterized and evaluated for anti-glycation activity. In vitro inhibition of advanced glycation end products (AGEs) by eugenol was found to be highest. Preliminary biophysical analysis and blind docking studies to understand eugenol-albumin interaction indicated eugenol to possess strong binding affinity for surface exposed lysines. However, binding of eugenol to bovine serum albumin (BSA) did not result in significant change in secondary structure of protein. In vivo diabetic mice model studies with eugenol showed reduction in blood glucose levels by 38% likely due to inhibition of alpha-glucosidase while insulin and glycated hemoglobin levels remain unchanged. Western blotting using anti-AGE antibody and mass spectrometry detected notably fewer AGE modified peptides upon eugenol treatment both in vivo and in vitro. Histopathological examination revealed comparatively lesser lesions in eugenol-treated mice. Thus, we propose eugenol has dual mode of action in combating diabetes; it lowers blood glucose by inhibiting a-glucosidase and prevents AGE formation by binding to epsilon-amine group on lysine, protecting it from glycation, offering potential use in diabetic management.

1 aSingh, Priyanka1 aJayaramaiah, Ramesha, H.1 aAgawane, Sachin, B.1 aVannuruswamy, Garikapati1 aKorwar, Arvind, M.1 aAnand, Atul1 aDhaygude, Vitthal, S.1 aShaikh, Mahemud, L.1 aJoshi, Rakesh, S.1 aBoppana, Ramanamurthy1 aKulkarni, Mahesh, J.1 aThulasiram, Hirekodathakallu, V.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/potential-dual-role-eugenol-inhibiting-advanced-glycation-end-products-diabetes-proteomic-002720nas a2200289 4500008004100000022001400041245015100055210006900206260008100275300002200356490000700378520163500385653001302020653001402033653001402047653002102061653001002082653002202092100003902114700002502153700002502178700002002203700002302223700002902246700002502275856013002300 2016 eng d a1542-641600aTargeted quantification of N-1-(carboxymethyl) valine and N-1-(carboxyethyl) valine peptides of beta-hemoglobin for better diagnostics in diabetes0 aTargeted quantification of N1carboxymethyl valine and N1carboxye a236 GRAYS INN RD, FLOOR 6, LONDON WC1X 8HL, ENGLANDbBIOMED CENTRAL LTDcMAR aArticle Number: 70 v133 a

Background: N-1-(Deoxyfructosyl) valine (DFV) beta-hemoglobin (beta-Hb), commonly referred as HbA1c, is widely used diagnostic marker in diabetes, believed to provide glycemic status of preceding 90-120 days. However, the turnover of hemoglobin is about 120 days, the DFV-beta-Hb, an early and reversible glycation product eventually may undergo irreversible advanced glycation modifications such as carboxymethylation or carboxyethylation. Hence quantification of N-1-(carboxymethyl) valine (CMV) and N-1-(carboxyethyl) valine (CEV) peptides of beta-Hb would be useful in assessing actual glycemic status. Results: Fragment ion library for synthetically glycated peptides of hemoglobin was generated by using high resolution-accurate mass spectrometry (HR/AM). Using parallel reaction monitoring, deoxyfructosylated, carboxymethylated and carboxyethylated peptides of hemoglobin were quantified in clinical samples from healthy control, pre-diabetes, diabetes and poorly controlled diabetes. For the first time, we report N-1-beta-valine undergoes carboxyethylation and mass spectrometric quantification of CMV and CEV peptides of beta-hemoglobin. Carboxymethylation was found to be the most abundant modification of N-1-beta-valine. Both CMV-beta-Hb and CEV-beta-Hb peptides showed better correlation with severity of diabetes in terms of fasting glucose, postprandial glucose and microalbuminuria. Conclusions: This study reports carboxymethylation as a predominant modification of N-1-beta-valine of Hb, and quantification of CMV-beta-Hb and CEV-beta-Hb could be useful parameter for assessing the severity of diabetes.

10aDiabetes10aDiagnosis10aGlycation10aGlycemic control10aHbA1c10aMass spectrometry1 aJagadeeshaprasad, Mashanipalya, G.1 aBatkulwar, Kedar, B.1 aMeshram, Nishita, N.1 aTiwari, Shalbha1 aKorwar, Arvind, M.1 aUnnikrishnan, Ambika, G.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/targeted-quantification-n-1-carboxymethyl-valine-and-n-1-carboxyethyl-valine-peptides-beta01499nas a2200145 4500008004100000245007600041210006900117300001200186520093800198100002901136700002301165700003901188700002501227856010101252 2017 eng d00aTargeted quantification of the glycated peptides of human serum albumin0 aTargeted quantification of the glycated peptides of human serum  a403-4163 a

Glycated human serum albumin (HSA) serves as an important marker for monitoring the glycemic status. Developing methods for unambiguous identification and quantification of glycated peptides of HSA using high-throughput technologies such as mass spectrometry has a great clinical significance. The following protocol describes the construction of reference spectral libraries for Amadori-modified lysine (AML), N(ε)-(carboxymethyl) lysine (CML)-, and N(ε)-(carboxyethyl)lysine (CEL)-modified peptides of synthetically modified HSA using high-resolution mass spectrometers. The protocol also describes work flows, for unambiguous identification and quantification of glycated modified peptides of HSA in clinical plasma using standard spectral libraries by various mass spectrometry approaches such as parallel reaction monitoring (PRM), sequential window acquisition of all theoretical fragment ion spectra (SWATH), and MSE.

1 aGarikapati, Vannuruswamy1 aKorwar, Arvind, M.1 aJagadeeshaprasad, Mashanipalya, G.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/targeted-quantification-glycated-peptides-human-serum-albumin02277nas a2200337 4500008004100000022001400041245011600055210006900171260000800240300001400248490000700262520120700269653000601476653000601482653002101488653001101509653003001520653002801550653003101578100001801609700001701627700002501644700002401669700001701693700002301710700002401733700002201757700002201779700001901801856011901820 2022 eng d a1381-199100aDetection of a target protein (GroEl2) in Mycobacterium tuberculosis using a derivative of 1,2,4-triazolethiols0 aDetection of a target protein GroEl2 in Mycobacterium tuberculos cOCT a2535-25480 v263 a

Herein, we identified a potent lead compound RRA2, within a series of 54 derivatives of 1,2,4-triazolethiols (exhibit good potency as an anti-mycobacterial agents) against intracellular Mycobacterium tuberculosis (Mtb). Compound RRA2 showed significant mycobactericidal activity against active stage Mycobacterium bovis BCG and Mtb with minimum inhibitory concentration (MIC) values of 2.3 and 2.0 mu g/mL, respectively. At MIC value, RRA2 compound yielded 0.82 log reduction of colony-forming unit (cfu) against non-replicating Mtb. Furthermore, RRA2 compound was selected for further target identification due to the presence of alkyne group, showing higher selectivity index (> 66.66 +/- 0.22, in non-replicating stage). Using ``click'' chemistry, we synthesized the biotin linker-RRA2 conjugate, purified with HPLC method and confirmed the conjugation of biotin linker-RRA2 complex by HR-MS analysis. Furthermore, we successfully pulled down and identified a specific target protein GroEl2, from Mtb whole-cell extract. Furthermore, computational molecular modeling indicated RRA2 could interact with GroEl2, which explains the structure-activity relationship observed in this study.

10a110a210a4-triazolethiols10aGroEl210aMycobactericidal activity10aMycobacterium bovis BCG10aMycobacterium tuberculosis1 aSarkar, Sampa1 aSwami, Sagar1 aSoni, Sarvesh, Kumar1 aHolien, Jessica, K.1 aKhan, Arshad1 aKorwar, Arvind, M.1 aLikhite, Anjali, P.1 aJoshi, Ramesh, A.1 aJoshi, Rohini, R.1 aSarkar, Dhiman uhttps://library.ncl.res.in/content/detection-target-protein-groel2-mycobacterium-tuberculosis-using-derivative-124