A Continuous Spectrophotometric and Fluorometric Assay for Protein Tyrosine Phosphatase Using Phosphotyrosine-Containing Peptides
dc.contributor.author | Zhang Z. Y. , | en_US |
dc.contributor.author | Maclean D. , | en_US |
dc.contributor.author | Thiemesefler A. M. , | en_US |
dc.contributor.author | Roeske R. W. , | en_US |
dc.contributor.author | Dixon J. E. , | en_US |
dc.date.accessioned | 2006-04-10T15:45:17Z | |
dc.date.available | 2006-04-10T15:45:17Z | |
dc.date.issued | 1993-05-15 | en_US |
dc.identifier.citation | Zhang Z. Y., , Maclean D., , Thiemesefler A. M., , Roeske R. W., , Dixon J. E., (1993/05/15)."A Continuous Spectrophotometric and Fluorometric Assay for Protein Tyrosine Phosphatase Using Phosphotyrosine-Containing Peptides." Analytical Biochemistry 211(1): 7-15. <http://hdl.handle.net/2027.42/30790> | en_US |
dc.identifier.uri | http://www.sciencedirect.com/science/article/B6W9V-45R7D3M-3C/2/92103ddbd66b3bf14eee5cc643d86e99 | en_US |
dc.identifier.uri | https://hdl.handle.net/2027.42/30790 | |
dc.identifier.uri | http://www.ncbi.nlm.nih.gov/sites/entrez?cmd=retrieve&db=pubmed&list_uids=7686722&dopt=citation | en_US |
dc.description.abstract | Two continuous assays for protein tyrosine phosphatases (PTPases) have been developed using phosphotyrosine containing peptide substrates. These assays are based on the marked differences in the spectra of the peptide before and after the removal of the phosphate group. The increase in the absorbance at 282 nm or the fluorescence at 305 nm of the peptide upon the action of PTPase can be followed continuously and the resulting progress curve (time course) can be analyzed directly using the integrated form of the Michaelis-Menten equation. The procedure is convenient and efficient, since both cat and Km values can be obtained in a single run. The difference absorption coefficient ([Delta]epsi) at 282 nm is relatively insensitive to the pH of the reaction media. These techniques were applied to two homogeneous recombinant PTPases employing six phosphotyrosine-containing peptides. Km and cat values obtained from the progress curve analysis were similar to those determined by the traditional initial rate inorganic phosphate assay. The peptides corresponding to autophosphorylation sites in Neu, p56lck, and p60src proteins show distinct behavior with the Yersinia PTPase, Yop51*, and the mammalian PTPase (PTP1U323). In both cases, the cat values were relatively constant for all the peptides tested whereas the Km values were very sensitive to the amino acid sequence surrounding the tyrosine residue, especially in the case of Yop51*. Thus, both Yop51* and PTP1U323 show differential recognition of the phosphotyrosyl residues in the context of distinct primary structure of peptide substrates. | en_US |
dc.format.extent | 759090 bytes | |
dc.format.extent | 3118 bytes | |
dc.format.mimetype | application/pdf | |
dc.format.mimetype | text/plain | |
dc.language.iso | en_US | |
dc.publisher | Elsevier | en_US |
dc.title | A Continuous Spectrophotometric and Fluorometric Assay for Protein Tyrosine Phosphatase Using Phosphotyrosine-Containing Peptides | en_US |
dc.type | Article | en_US |
dc.rights.robots | IndexNoFollow | en_US |
dc.subject.hlbsecondlevel | Public Health | en_US |
dc.subject.hlbsecondlevel | Chemistry | en_US |
dc.subject.hlbsecondlevel | Chemical Engineering | en_US |
dc.subject.hlbsecondlevel | Biological Chemistry | en_US |
dc.subject.hlbtoplevel | Engineering | en_US |
dc.subject.hlbtoplevel | Science | en_US |
dc.subject.hlbtoplevel | Health Sciences | en_US |
dc.description.peerreviewed | Peer Reviewed | en_US |
dc.contributor.affiliationum | Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, MI, USA; Walther Cancer Institute, Ann Arbor, MI, USA. | en_US |
dc.contributor.affiliationum | Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, MI, USA; Walther Cancer Institute, Ann Arbor, MI, USA. | en_US |
dc.contributor.affiliationother | Parke-Davis Research Divsion, Warner Lambert Company, Ann Arbor, MI, USA. | en_US |
dc.contributor.affiliationother | Parke-Davis Research Divsion, Warner Lambert Company, Ann Arbor, MI, USA. | en_US |
dc.contributor.affiliationother | Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis, Indiana, USA. | en_US |
dc.identifier.pmid | 7686722 | en_US |
dc.description.bitstreamurl | http://deepblue.lib.umich.edu/bitstream/2027.42/30790/1/0000444.pdf | en_US |
dc.identifier.doi | http://dx.doi.org/10.1006/abio.1993.1224 | en_US |
dc.identifier.source | Analytical Biochemistry | en_US |
dc.owningcollname | Interdisciplinary and Peer-Reviewed |
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