0000036364 00000 n Control of protein immobilization: Coupling immobilization and site-directed mutagenesis to improve biocatalyst or biosensor performance. 0000024490 00000 n Determination of Peptide and Protein Disulfide Linkages by MALDI Mass Spectrometry. Copyright © 1992 Published by Elsevier Inc. https://doi.org/10.1016/0003-2697(92)90013-W. Journal of the American Society for Mass Spectrometry. 151 0 obj 10.1002/1096-9888(200009)35:9<1148::AID-JMS982>3.0.CO;2-R. Assessment of the kinetics and sites of reaction of some Immobiline chemicals with proteins and peptides by matrix‐assisted laser desorption/ionisation time‐of‐flight mass spectrometry.
Use the link below to share a full-text version of this article with your friends and colleagues. /Size 153 It is recognised that gel‐separated proteins can experience a frequent modification provoked by the interaction of unpolymerized acrylamide monomers with the thiol group of cysteine to form a β‐cysteinyl‐S‐propionamide adduct. 0000026080 00000 n trailer Number of times cited according to CrossRef: Nanomaterial-Immobilized Biocatalysts for Biofuel Production from Lignocellulose Biomass. 94 0 obj Structure-Toxicity Analysis of Type-2 Alkenes: In Vitro Neurotoxicity. These high resolution data suggest that, for short incubation times (≥1 hour) and in the absence of cysteine, the ϵ‐NH2 group of lysine is the likely adduction site of acrylamide. /N 5 0000013485 00000 n stream endobj [95 0 R 0000024815 00000 n 0000004185 00000 n 0000002734 00000 n /MediaBox[0 0 595 791] 0000027979 00000 n and you may need to create a new Wiley Online Library account. 92 0 obj /T 194758 Chemical Reagents for Protein Modification, Fourth Edition. Acrylamide mainly interacted with the thiol (-SH) residue of CK-BB and resulted in alkylation. endobj xref 0000003151 00000 n Working off-campus? 0000002817 00000 n 0000028731 00000 n 10.1002/1522-2683(200011)21:17<3684::AID-ELPS3684>3.0.CO;2-E, Rapid Communications in Mass Spectrometry, https://doi.org/10.1002/(SICI)1097-0231(20000530)14:10<840::AID-RCM953>3.0.CO;2-M. Enter your email address below and we will send you your username, If the address matches an existing account you will receive an email with instructions to retrieve your username, By continuing to browse this site, you agree to its use of cookies as described in our, I have read and accept the Wiley Online Library Terms and Conditions of Use.
0000029155 00000 n Investigating the reaction of a number of gel electrophoresis cross‐linkers with β‐lactoglobulin by matrix assisted laser desorption/ionization‐ mass spectrometry. 0000013950 00000 n 0000023142 00000 n
Investigating the reaction of a novel silica capillary coating compound with proteins/peptides by matrix‐assisted laser desorption/ionisation time‐of‐flight mass spectrometry. To emphasise this conclusion we have used matrix‐assisted laser desorption/ionisation with a reflectron time‐of‐flight mass spectrometer to examine a number of cysteine‐free proteins incubated for various intervals with 30 mM acrylamide monomers at pH 9.5. Acceleration and Improvement of Protein Identification by Mass Spectrometry.
Protein alkylation in the presence/absence of thiourea in proteome analysis: A matrix assisted laser desorption/ionization‐time of flight‐mass spectrometry investigation. Longer incubation times (≥24 hours) with acrylamide monomers rendered the role of Cys as the favourite alkylation site less evident. %PDF-1.3 0000002571 00000 n 0000001550 00000 n The monomeric form is widely … <> If you do not receive an email within 10 minutes, your email address may not be registered, endobj 0000026875 00000 n
Protein alkylation by acrylamide, its ‐substituted derivatives and cross‐linkers and its relevance to proteomics: A matrix assisted laser desorption/ionization‐time of flight‐mass spectrometry study. Qc�c>�������
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[�m�|�@v�?ӝ'@��a��2!L��4e��Wf���#�>�i��R V�u� Reactivity of the NS2/3(907–1206)ASK4 protein with β‐mercaptoethanol studied by electrospray ion trap mass spectrometry. A new method for analysis of disulfide-containing proteins by matrix-assisted laser desorption ionization (MALDI) mass spectrometry. The full text of this article hosted at iucr.org is unavailable due to technical difficulties. 0000023282 00000 n 0000026652 00000 n 147 0 R Copyright © 2000 John Wiley & Sons, Ltd. Cysteine in proteins is easily alkylated by reaction of proteins in aqueous solution with acrylamide. PTH-Cys-S-Pam was first noticed during sequencing polypeptides blotted onto polyvinylidene difluoride membranes from polyacrylamide gels, in which cysteine had reacted with residual unpolymerized acrylamide.
95 0 obj Its phenylthiohydantoin derivative, PTH-Cys-S-Pam, is easily separated from other PTH-amino acids by HPLC and is thus useful for cysteine identification during protein sequencing. ] %����
0000001733 00000 n The work described here was partially supported by the Arizona State University Center for the Study of Early Events in Photosynthesis. >> Alkylation of cysteine in proteins with acrylamide under mildly alkaline conditions yields a thioether derivative, Cys-S-β-propionamide (Cys-S-Pam), which is stable during automated Edman degradation. 0000013296 00000 n /Prev 194749 0000025926 00000 n The Center is funded by U.S. Department of Energy Grant DE-FG-88-ER13969 as a part of the USDA/DOE/NSF Plant Science Centers Program. 93 0 obj 149 0 R Proteomic analysis of acrylamide-protein adduct formation in rat brain synaptosomes. 0000052719 00000 n 0000060458 00000 n /E 65218 148 0 R /Contents [143 0 R 0000000015 00000 n 0000026361 00000 n
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0000026943 00000 n ProteoSys AG, … 0000003068 00000 n 10.1002/1522-2683(200105)22:9<1633::AID-ELPS1633>3.0.CO;2-C. The protein sequencer was obtained with funds from NSF Grant BBS88-04992 (NSF Biological Centers Program). /Type/Page 97 0 obj 0000002652 00000 n endobj 0000027675 00000 n 0000002191 00000 n Analysis of relative isotopologue abundances for quantitative profiling of complex protein mixtures labelled with the acrylamide/D 3 ‐acrylamide alkylation tag system Michael A. Cahill. /Info 87 0 R /Resources 105 0 R Modification of Sulfur-Containing Amino Acids in Proteins. endobj 0000039115 00000 n /L 196642 >>
We achieved best results with acrylamide as alkylation reagent, whereas the highest numbers of peptide spectral matches were obtained when reducing with dithiothreitol and β-mercaptoethanol for the in … 0000058938 00000 n 10.1002/1097-0231(20010215)15:3<210::AID-RCM215>3.0.CO;2-5. << 0000001677 00000 n
96 0 obj 0000023555 00000 n 0000026140 00000 n Protein Adduct Formation as a Molecular Mechanism in Neurotoxicity. These high resolution data suggest that, for short incubation times (≥1 hour) and in the absence of cysteine, the ϵ‐NH 2 group of lysine is the likely adduction site of acrylamide. Rapid Communications in Mass Spectrometry. endobj 0000061986 00000 n 0000025157 00000 n 0000025630 00000 n 0000025092 00000 n 145 0 R Concealment of epitope by reduction and alkylation in prion protein.
0000054852 00000 n Preparative isolation of protein complexes and other bioparticles by elution from polyacrylamide gels. 366 /F 97 0 R <. 0000002212 00000 n 98 0 obj %%EOF While cysteine alkylation with acrylamide is an undesired reaction that frequently occurs during polyacrylamide gel electrophoresis,11 there are several properties that make acryl-amide a very useful … startxref /Parent 86 0 R /O 96 A computational docking simulation supported that acrylamide directly bound to the active site of CK-BB … Learn about our remote access options, GlaxoWellcome Medicines Research Centre, via Fleming 4, Verona, Italy, E-mail address: mh12706@Glaxo Wellcome.co.uk, University of Verona, Department of Agricultural and Industrial Biotechnologies strada le Grazie 15, Cà Vignal, Verona, Italy. 0000002985 00000 n 10.1002/1522-2683(200106)22:10<2066::AID-ELPS2066>3.0.CO;2-8. >A�O���@
��820�� ��@� Keywords: acrylamide; alkylation reactions INTRODUCTION Acrylamide (AA), an industrially produced reactive molecule, is used worldwide to synthesize polyacrylamide. 0000036307 00000 n /Rotate 0
It has the same HPLC retention time as the product formed from cysteine when sequencing proteins alkylated with acrylamide.