Reference: Todd RJ, et al. (1994)
Reference Help
Abstract
Mechanisms of protein retention in immobilized metal-affinity chromatography (IMAC) have been probed using a set of Saccharomyces cerevisiae iso-1-cytochrome c histidine variants constructed by site-directed mutagenesis. Proteins containing a single accessible histidine exhibit Langmuir-type isotherms with maximum protein binding capacities between 5 and 10% of the maximum copper loading and the capacity of the support to bind imidazole. A simple model that assumes that the copper sites are densely packed and can be blocked by protein adsorption yields binding constants for single-histidine proteins that are similar to the binding constant for free imidazole. Proteins containing multiple accessible histidines do not exhibit simple Langmuir-type behavior; they appear to interact with the support by simultaneous coordination to more than one metal ion, the result of which is to increase the apparent binding affinity by as much as a factor of 1000. The protein binding constant depends on the availability of copper sites: binding is significantly weaker at low surface concentrations of copper that presumably cannot support multiple-site interactions. The protein binding capacity drops to zero at copper loadings less than one-half the maximum, indicating that immobilized iminodiacetic acid ligands are sufficiently close together that two can coordinate a single copper ion, which precludes its interaction with a protein. Protein adsorption via multiple-site coordination has important consequences for the optimization of IMAC separations and the design of new IMAC supports.
- Reference Type
-
Journal Article |
Research Support, Non-U.S. Gov't |
Research Support, U.S. Gov't, Non-P.H.S. |
Research Support, U.S. Gov't, P.H.S.
- Authors
-
Todd RJ,
Johnson RD,
Arnold FH
... Show all
Show fewer
- CYC1
Gene Ontology Annotations
Increase the total number of rows showing on this page using the pull-down located below the table, or use the page
scroll at the table's top right to browse through the table's pages; use the arrows to the right of a column header
to sort by that column; filter the table using the "Filter" box at the top of the table.
Evidence ID |
Analyze ID |
Gene/Complex |
Systematic Name/Complex Accession |
Qualifier |
Gene Ontology Term ID |
Gene Ontology Term |
Aspect |
Annotation Extension |
Evidence |
Method |
Source |
Assigned On |
Reference |
Phenotype Annotations
Increase the total number of rows showing on this page using the pull-down located below the table, or use the page
scroll at the table's top right to browse through the table's pages; use the arrows to the right of a column header
to sort by that column; filter the table using the "Filter" box at the top of the table; click on the small "i"
buttons located within a cell for an annotation to view further details.
Evidence ID |
Analyze ID |
Gene |
Gene Systematic Name |
Phenotype |
Experiment Type |
Experiment Type Category |
Mutant Information |
Strain Background |
Chemical |
Details |
Reference |
Disease Annotations
Increase the total number of rows showing on this page using the pull-down located below the table, or use the page
scroll at the table's top right to browse through the table's pages; use the arrows to the right of a column header
to sort by that column; filter the table using the "Filter" box at the top of the table.
Evidence ID |
Analyze ID |
Gene |
Gene Systematic Name |
Disease Ontology Term |
Disease Ontology Term ID |
Qualifier |
Evidence |
Method |
Source |
Assigned On |
|
Reference |
Regulation Annotations
Increase the total number of rows displayed on this page using the pull-down located below the table, or use the
page scroll at the table's top right to browse through the table's pages; use the arrows to the right of a column
header to sort by that column; to filter the table by a specific experiment type, type a keyword into the Filter box
(for example, “microarray”); download this table as a .txt file using the Download button or click Analyze to
further view and analyze the list of target genes using GO Term Finder, GO Slim Mapper, or SPELL.
Evidence ID |
Analyze ID |
Regulator |
Regulator Systematic Name |
Target |
Target Systematic Name |
Direction |
Regulation of |
Happens During |
Regulator Type |
Direction |
Regulation Of |
Happens During |
Method |
Evidence |
Strain Background |
Reference |
Post-translational Modifications
Increase the total number of rows showing on this page by using the pull-down located below the table, or use the
page scroll at the table's top right to browse through its pages; use the arrows to the right of a column header to
sort by that column; filter the table using the "Filter" box at the top of the table.
|
|
|
|
Site |
|
Modification |
Modifier |
Source |
Reference |
Interaction Annotations
Genetic Interactions
Increase the total number of rows showing on this page by using the pull-down located below the table, or use the
page scroll at the table's top right to browse through the table's pages; use the arrows to the right of a column
header to sort by that column; filter the table using the "Filter" box at the top of the table; click on the small
"i" buttons located within a cell for an annotation to view further details about experiment type and any other
genes involved in the interaction.
Evidence ID |
Analyze ID |
|
Interactor |
Interactor Systematic Name |
Interactor |
Interactor Systematic Name |
Allele |
Assay |
Annotation |
Action |
Phenotype |
SGA score |
P-value |
Source |
Reference |
Note |
Physical Interactions
Increase the total number of rows showing on this page by using the pull-down located below the table, or use the
page scroll at the table's top right to browse through the table's pages; use the arrows to the right of a column
header to sort by that column; filter the table using the "Filter" box at the top of the table; click on the small
"i" buttons located within a cell for an annotation to view further details about experiment type and any other
genes involved in the interaction.
Evidence ID |
Analyze ID |
|
Interactor |
Interactor Systematic Name |
Interactor |
Interactor Systematic Name |
Assay |
Annotation |
Action |
Modification |
Source |
Reference |
Note |
Functional Complementation Annotations
Increase the total number of rows showing on this page by using the pull-down located below the table, or use the
page scroll at the table's top right to browse through its pages; use the arrows to the right of a column header to
sort by that column; filter the table using the "Filter" box at the top of the table.
Complement ID |
Locus ID |
Gene |
Species |
Gene ID |
Strain background |
Direction |
Details |
Source |
Reference |