Show simple item record

Isoelectric focusing nonporous silica reversed-phase high-performance liquid chromatography/electrospray ionization time-of-flight mass spectrometry: a three-dimensional liquid-phase protein separation method as applied to the human erythroleukemia cell-line

dc.contributor.authorWall, Daniel B.en_US
dc.contributor.authorKachman, Maureen T.en_US
dc.contributor.authorGong, Siyuan S.en_US
dc.contributor.authorParus, Stephan J.en_US
dc.contributor.authorLong, Michael W.en_US
dc.contributor.authorLubman, David M.en_US
dc.date.accessioned2006-04-19T14:08:10Z
dc.date.available2006-04-19T14:08:10Z
dc.date.issued2001-09-30en_US
dc.identifier.citationWall, Daniel B.; Kachman, Maureen T.; Gong, Siyuan S.; Parus, Stephen J.; Long, Michael W.; Lubman, David M. (2001)."Isoelectric focusing nonporous silica reversed-phase high-performance liquid chromatography/electrospray ionization time-of-flight mass spectrometry: a three-dimensional liquid-phase protein separation method as applied to the human erythroleukemia cell-line." Rapid Communications in Mass Spectrometry 15(18): 1649-1661. <http://hdl.handle.net/2027.42/35080>en_US
dc.identifier.issn0951-4198en_US
dc.identifier.issn1097-0231en_US
dc.identifier.urihttps://hdl.handle.net/2027.42/35080
dc.identifier.urihttp://www.ncbi.nlm.nih.gov/sites/entrez?cmd=retrieve&db=pubmed&list_uids=11555863&dopt=citationen_US
dc.description.abstractA liquid-phase three-dimensional protein separation method has been developed that is used to separate the cytosolic fraction of a HEL cell lysate via isoelectric focusing (IEF), nonporous silica (NPS) reversed-phase high-performance liquid chromatography (RP-HPLC) and electrospray ionization time-of-flight mass spectrometry (ESI-TOFMS), respectively. Several hundred unique protein molecular weights were observed in a pI range from 4.8 to 8.5 and a mass range from 5 to 85 14kDa. Proteins were positively identified by analysis of the pI (±0.5 pI units), an intact protein molecular weight (±150 14ppm), and peptide mass mapping results. Using the molecular weight (MW) and peptide mapping results of identified proteins it was possible to characterize their posttranslational (PTMs) and/or sequence modifications. PTMs were detected on both forms of cytosolic actin, heat shock 90 Β, HINT and Α-enolase. Sequence modifications or conflicts were observed for Β-and Γ-actin, ATP Β-synthase and heat shock 90 Β. IEF-NPS-RP-HPLC/ESI-TOFMS was used to determine experimental pI, MW and relative hydrophobicity values for each protein detected. This data was used to generate a 2-D pI-MS protein map, where proteins are displayed according to their pI and molecular weight. Protein molecular weight peaks are represented as bands in the 2-D pI-MS image where the gray scale of each band is proportional to the intensity of the protein molecular weight peak. In addition, a third hydrophobicity dimension (%B) was added as the % acetonitrile elution to generate a 3-D pI-MS-%B plot where each protein can be tagged according to three parameters. Copyright © 2001 John Wiley & Sons, Ltd.en_US
dc.format.extent503942 bytes
dc.format.extent3118 bytes
dc.format.mimetypeapplication/pdf
dc.format.mimetypetext/plain
dc.language.isoen_US
dc.publisherJohn Wiley & Sons, Ltd.en_US
dc.subject.otherChemistryen_US
dc.subject.otherAnalytical Chemistry and Spectroscopyen_US
dc.titleIsoelectric focusing nonporous silica reversed-phase high-performance liquid chromatography/electrospray ionization time-of-flight mass spectrometry: a three-dimensional liquid-phase protein separation method as applied to the human erythroleukemia cell-lineen_US
dc.typeArticleen_US
dc.rights.robotsIndexNoFollowen_US
dc.subject.hlbtoplevelScienceen_US
dc.description.peerreviewedPeer Revieweden_US
dc.contributor.affiliationumDepartment of Chemistry, The University of Michigan, Ann Arbor, MI 48109-1055, USAen_US
dc.contributor.affiliationumDepartment of Chemistry, The University of Michigan, Ann Arbor, MI 48109-1055, USAen_US
dc.contributor.affiliationumDepartment of Chemistry, The University of Michigan, Ann Arbor, MI 48109-1055, USAen_US
dc.contributor.affiliationumDepartment of Chemistry, The University of Michigan, Ann Arbor, MI 48109-1055, USAen_US
dc.contributor.affiliationumDepartment of Pediatrics, School of Medicine, The University of Michigan, Ann Arbor, MI 48109, USAen_US
dc.contributor.affiliationumDepartment of Chemistry, The University of Michigan, Ann Arbor, MI 48109-1055, USA ; Department of Chemistry, The University of Michigan, Ann Arbor, MI 48109-1055, USA.en_US
dc.identifier.pmid11555863en_US
dc.description.bitstreamurlhttp://deepblue.lib.umich.edu/bitstream/2027.42/35080/1/421_ftp.pdfen_US
dc.identifier.doihttp://dx.doi.org/10.1002/rcm.421en_US
dc.identifier.sourceRapid Communications in Mass Spectrometryen_US
dc.owningcollnameInterdisciplinary and Peer-Reviewed


Files in this item

Show simple item record

Remediation of Harmful Language

The University of Michigan Library aims to describe library materials in a way that respects the people and communities who create, use, and are represented in our collections. Report harmful or offensive language in catalog records, finding aids, or elsewhere in our collections anonymously through our metadata feedback form. More information at Remediation of Harmful Language.

Accessibility

If you are unable to use this file in its current format, please select the Contact Us link and we can modify it to make it more accessible to you.