MALDI-TOF High Mass Calibration up to 200 kDa Using Human Recombinant 16 kDa Protein Histidine Phosphatase Aggregates
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{"title"=>"MALDI-TOF high mass calibration up to 200 kDa using human recombinant 16 kDa protein histidine phosphatase aggregates", "type"=>"journal", "authors"=>[{"first_name"=>"Katrin", "last_name"=>"Ludwig", "scopus_author_id"=>"35240565200"}, {"first_name"=>"Schähdi", "last_name"=>"Habbach", "scopus_author_id"=>"47861846500"}, {"first_name"=>"Josef", "last_name"=>"Krieglstein", "scopus_author_id"=>"55110761500"}, {"first_name"=>"Susanne", "last_name"=>"Klumpp", "scopus_author_id"=>"7005580926"}, {"first_name"=>"Simone", "last_name"=>"König", "scopus_author_id"=>"7101897399"}], "year"=>2011, "source"=>"PLoS ONE", "identifiers"=>{"pui"=>"362371984", "sgr"=>"80051839027", "issn"=>"19326203", "pmid"=>"21876758", "scopus"=>"2-s2.0-80051839027", "doi"=>"10.1371/journal.pone.0023612"}, "id"=>"0a78820c-46d2-3503-b115-52faf95aaacc", "abstract"=>"Protein histidine phosphatase (PHP) is an enzyme which removes phosphate groups from histidine residues. It was described for vertebrates in the year 2002. The recombinant human 16 kDa protein forms multimeric complexes in physiological buffer and in the gas phase. High-mass calibration in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has remained a problem due to the lack of suitable standards. Large proteins can hardly be freed of their substructural microheterogeneity by classical purification procedures so that their use as calibrants is limited. A small adduct-forming protein of validated quality is a valuable alternative for that purpose.", "link"=>"http://www.mendeley.com/research/malditof-high-mass-calibration-up-200-kda-using-human-recombinant-16-kda-protein-histidine-phosphata", "reader_count"=>17, "reader_count_by_academic_status"=>{"Researcher"=>7, "Student > Ph. D. Student"=>5, "Other"=>1, "Student > Master"=>1, "Student > Bachelor"=>2, "Professor"=>1}, "reader_count_by_user_role"=>{"Researcher"=>7, "Student > Ph. D. Student"=>5, "Other"=>1, "Student > Master"=>1, "Student > Bachelor"=>2, "Professor"=>1}, "reader_count_by_subject_area"=>{"Engineering"=>1, "Biochemistry, Genetics and Molecular Biology"=>2, "Agricultural and Biological Sciences"=>8, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Physics and Astronomy"=>1, "Chemistry"=>3, "Computer Science"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Chemistry"=>{"Chemistry"=>3}, "Physics and Astronomy"=>{"Physics and Astronomy"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>8}, "Computer Science"=>{"Computer Science"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}}, "reader_count_by_country"=>{"Germany"=>1}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/744817"], "description"=>"<p>Using <b>A</b>) Superdex 200 (Signal d was outside the separation range of the column so that cluster size was estimated.) and <b>B</b>) native gel electrophoresis (several multimer signals, but only one monomer band).</p>", "links"=>[], "tags"=>["php", "aggregate"], "article_id"=>415195, "categories"=>["Physics", "Biochemistry", "Chemistry", "Biophysics"], "users"=>["Katrin Ludwig", "Schähdi Habbach", "Josef Krieglstein", "Susanne Klumpp", "Simone König"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0023612.g003", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Determination_of_PHP_aggregate_sizes_/415195", "title"=>"Determination of PHP aggregate sizes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-08-18 01:26:35"}
  • {"files"=>["https://ndownloader.figshare.com/files/375335", "https://ndownloader.figshare.com/files/375365", "https://ndownloader.figshare.com/files/375384", "https://ndownloader.figshare.com/files/375410", "https://ndownloader.figshare.com/files/375426", "https://ndownloader.figshare.com/files/375454", "https://ndownloader.figshare.com/files/375482", "https://ndownloader.figshare.com/files/375512"], "description"=>"<div><h3>Background</h3><p>Protein histidine phosphatase (PHP) is an enzyme which removes phosphate groups from histidine residues. It was described for vertebrates in the year 2002. The recombinant human 16 kDa protein forms multimeric complexes in physiological buffer and in the gas phase. High-mass calibration in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has remained a problem due to the lack of suitable standards. Large proteins can hardly be freed of their substructural microheterogeneity by classical purification procedures so that their use as calibrants is limited. A small adduct-forming protein of validated quality is a valuable alternative for that purpose.</p> <h3>Methodology/Principal Findings</h3><p>Three major PHP clusters of ∼113, 209 and >600 kDa were observed in gel filtration analysis. Re-chromatography of the monomer peak showed the same cluster distribution. The tendency to associate was detected also in MALDI-TOF MS measuring regular adducts up to 200 kDa.</p> <h3>Conclusions/Significance</h3><p>PHP forms multimers consisting of up to more than 35 protein molecules. In MALDI-TOF MS it generates adduct ions every 16 kDa. The protein can be produced with high quality so that its use as calibration compound for high mass ranges above 100 kDa, where standards are difficult to obtain, is feasible.</p> </div>", "links"=>[], "tags"=>["maldi-tof", "calibration", "recombinant", "16", "kda", "histidine", "phosphatase", "aggregates"], "article_id"=>134124, "categories"=>["Physics", "Biochemistry", "Chemistry", "Biophysics"], "users"=>["Katrin Ludwig", "Schähdi Habbach", "Josef Krieglstein", "Susanne Klumpp", "Simone König"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0023612.s001", "https://dx.doi.org/10.1371/journal.pone.0023612.s002", "https://dx.doi.org/10.1371/journal.pone.0023612.s003", "https://dx.doi.org/10.1371/journal.pone.0023612.s004", "https://dx.doi.org/10.1371/journal.pone.0023612.s005", "https://dx.doi.org/10.1371/journal.pone.0023612.s006", "https://dx.doi.org/10.1371/journal.pone.0023612.s007", "https://dx.doi.org/10.1371/journal.pone.0023612.s008"], "stats"=>{"downloads"=>35, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/MALDI_TOF_High_Mass_Calibration_up_to_200_kDa_Using_Human_Recombinant_16_kDa_Protein_Histidine_Phosphatase_Aggregates/134124", "title"=>"MALDI-TOF High Mass Calibration up to 200 kDa Using Human Recombinant 16 kDa Protein Histidine Phosphatase Aggregates", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2011-08-18 01:08:44"}
  • {"files"=>["https://ndownloader.figshare.com/files/744934"], "description"=>"<p>Satellite peaks correspond to residual contaminants from protein isolation, which cannot be easily removed. PHP sequence is given as inset.</p>", "links"=>[], "tags"=>["nanospray", "desalted"], "article_id"=>415305, "categories"=>["Physics", "Biochemistry", "Chemistry", "Biophysics"], "users"=>["Katrin Ludwig", "Schähdi Habbach", "Josef Krieglstein", "Susanne Klumpp", "Simone König"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0023612.g004", "stats"=>{"downloads"=>1, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Q_TOF_nanospray_mass_spectrum_of_desalted_PHP_/415305", "title"=>"Q-TOF nanospray mass spectrum of desalted PHP.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-08-18 01:28:25"}
  • {"files"=>["https://ndownloader.figshare.com/files/744757"], "description"=>"<p>(<a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0023612#pone-0023612-g001\" target=\"_blank\">Figure 1</a>; a) sample 1, b/c) sample 2, d) sample 3) with respect to PHP <b>C</b>) activity (dephosphorylation of ACL) and <b>A/B</b>) purity. 1D-PAGE using PHP as control. <b>A</b>) Coomassie-staining. <b>B</b>) Detection with polyclonal PHP antibody.</p>", "links"=>[], "tags"=>["gel", "chromatography"], "article_id"=>415130, "categories"=>["Physics", "Biochemistry", "Chemistry", "Biophysics"], "users"=>["Katrin Ludwig", "Schähdi Habbach", "Josef Krieglstein", "Susanne Klumpp", "Simone König"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0023612.g002", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Quality_control_of_gel_chromatography_fractions_/415130", "title"=>"Quality control of gel chromatography fractions.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-08-18 01:25:30"}
  • {"files"=>["https://ndownloader.figshare.com/files/744689"], "description"=>"<p>(Superdex 75, 4°C, 280 nm). a) Monomer. Fractions b) and c) were pooled for further processing (see <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0023612#pone-0023612-g002\" target=\"_blank\">Figure 2</a>).</p>", "links"=>[], "tags"=>["gel", "filtration"], "article_id"=>415063, "categories"=>["Physics", "Biochemistry", "Chemistry", "Biophysics"], "users"=>["Katrin Ludwig", "Schähdi Habbach", "Josef Krieglstein", "Susanne Klumpp", "Simone König"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0023612.g001", "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_PHP_gel_filtration_profile_/415063", "title"=>"PHP gel filtration profile.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-08-18 01:24:23"}
  • {"files"=>["https://ndownloader.figshare.com/files/745080"], "description"=>"<p>Average molecular weights of singly and doubly protonated PHP multimers.</p>", "links"=>[], "tags"=>["molecular", "weights", "singly", "doubly", "protonated", "php"], "article_id"=>415446, "categories"=>["Physics", "Biochemistry", "Chemistry", "Biophysics"], "users"=>["Katrin Ludwig", "Schähdi Habbach", "Josef Krieglstein", "Susanne Klumpp", "Simone König"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0023612.t001", "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Average_molecular_weights_of_singly_and_doubly_protonated_PHP_multimers_/415446", "title"=>"Average molecular weights of singly and doubly protonated PHP multimers.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2011-08-18 01:30:46"}
  • {"files"=>["https://ndownloader.figshare.com/files/745020"], "description"=>"<p>Top: Monomer up to hexamer (observe zoom for range ><i>m/z</i> 3700); Below: Trimer to dodecamer. For peak assignment see <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0023612#pone-0023612-t001\" target=\"_blank\">Table 1</a>.</p>", "links"=>[], "tags"=>["spectra", "his-tagged"], "article_id"=>415393, "categories"=>["Physics", "Biochemistry", "Chemistry", "Biophysics"], "users"=>["Katrin Ludwig", "Schähdi Habbach", "Josef Krieglstein", "Susanne Klumpp", "Simone König"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0023612.g005", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_MALDI_TOF_spectra_of_His_tagged_PHP_/415393", "title"=>"MALDI-TOF spectra of His-tagged PHP.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-08-18 01:29:53"}

PMC Usage Stats | Further Information

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Relative Metric

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