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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
7
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pubmed:dateCreated |
1990-5-4
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pubmed:abstractText |
The divalent cation-binding properties of the human insulin receptor tyrosine kinase domain were examined kinetically and by electron paramagnetic resonance and circular dichroic spectroscopy. The protein-tyrosine kinase activity of the purified cytoplasmic domain can be activated nearly 10-fold by 3 mM Mn2+ in the presence or absence of 5 mM Mg2+. Electron paramagnetic resonance spectra of the purified, acid-denatured kinase domain and assays of EDTA-treated kinase show that the purified protein does not possess residual, tightly bound Mn2+. Electron paramagnetic resonance spectroscopy was used to directly measure the binding constant of the kinase domain for Mn2+. The results indicate that the recombinant cytoplasmic domain of the human insulin receptor does not bind Mn2+ tightly in the absence or presence of MgATP (Kd greater than 0.8 mM). Furthermore, the enzyme does not show a strong preference for MnATP binding when both MgATP and MnATP are present. The far-ultraviolet circular dichroic spectrum of this domain is characterized by a negative maximum at 207 nm. In the presence of Mn2+, but not Mg2+, changes in the mean residue-weight ellipticity at 207 nm occur that are consistent with a decrease in alpha-helical content. The addition of ATP to Mn2(+)-bound protein does not further perturb the spectrum. We conclude that Mn2+ ions, although they bind weakly, induce an activating conformational change in the secondary structure of the human insulin receptor cytoplasmic domain. Activation by Mn2+ is unlikely to be significant in intact cells, but it may mimic the action of a physiological activator.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/2157215-193671,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2157215-2442814,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2157215-2542339,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2157215-2544997,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2157215-2554291,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2157215-2833165,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2157215-3004334,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2157215-3100537,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2157215-3101064,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2157215-3360145,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2157215-4337505,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2157215-5432063,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2157215-6203118,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2157215-6286611,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2157215-6294652,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2157215-6325418,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2157215-6336757,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2157215-6368536,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2157215-6757253
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Apr
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pubmed:issn |
0027-8424
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
87
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
2805-9
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pubmed:dateRevised |
2009-11-19
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pubmed:meshHeading |
pubmed-meshheading:2157215-Circular Dichroism,
pubmed-meshheading:2157215-Electron Spin Resonance Spectroscopy,
pubmed-meshheading:2157215-Histones,
pubmed-meshheading:2157215-Humans,
pubmed-meshheading:2157215-Kinetics,
pubmed-meshheading:2157215-Manganese,
pubmed-meshheading:2157215-Protein Binding,
pubmed-meshheading:2157215-Protein Conformation,
pubmed-meshheading:2157215-Protein-Tyrosine Kinases,
pubmed-meshheading:2157215-Receptor, Insulin,
pubmed-meshheading:2157215-Recombinant Proteins,
pubmed-meshheading:2157215-Spectrophotometry, Ultraviolet
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pubmed:year |
1990
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pubmed:articleTitle |
Mn2(+)-binding properties of a recombinant protein-tyrosine kinase derived from the human insulin receptor.
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pubmed:affiliation |
Memorial Sloan-Kettering Cancer Center, New York, NY 10021.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
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