Ca 2+ -binding proteins: A comparative study of their behavior during high-performance liquid chromatography using gradient elution on reverse-phase supports
Analytical Biochemistry, ISSN: 0003-2697, Vol: 129, Issue: 1, Page: 120-131
1983
- 29Citations
- 1Captures
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Metrics Details
- Citations29
- Citation Indexes29
- 29
- CrossRef26
- Captures1
- Readers1
Article Description
Reverse-phase high-performance liquid chromatography has been shown to be applicable to the isolation of Ca 2+ -binding proteins, specifically parvalbumins, from tissue extracts or from preparations first purified by “conventional” chromatography. Through an investigation of the behavior of a series of Ca 2+ -binding proteins as a function of buffer composition, pH, and organic eluant it has been possible to define mild conditions allowing for chromatography of the proteins in their native states. The elution positions of parvalbumins were not observed to correlate with the “overall” protein hydrophobicity, calculated using hydrophobicity values for the individual amino acids, thus indicating that factors such as hydrophobic/hydrophilic surface areas are important in determining the degree of association with the support. The usefulness of reverse-phase chromatography as an analytical tool for determining protein homogeneity is illustrated. Samples which had been isolated via “conventional” chromatography methods, and thought to be homogeneous, were observed to contain multiple species of the same protein.
Bibliographic Details
http://www.sciencedirect.com/science/article/pii/000326978390060X; http://dx.doi.org/10.1016/0003-2697(83)90060-x; http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=0021103418&origin=inward; http://www.ncbi.nlm.nih.gov/pubmed/6859517; https://linkinghub.elsevier.com/retrieve/pii/000326978390060X; http://linkinghub.elsevier.com/retrieve/pii/000326978390060X; http://api.elsevier.com/content/article/PII:000326978390060X?httpAccept=text/xml; http://api.elsevier.com/content/article/PII:000326978390060X?httpAccept=text/plain; http://dx.doi.org/10.1016/0003-2697%2883%2990060-x; https://dx.doi.org/10.1016/0003-2697%2883%2990060-x
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