ISSN:
1612-1112
Keywords:
Discontinuous reversed-phase HPLC
;
Ribosomal proteins of archaebacteriumSulfolobus acidocaldarius
;
Load capacity for proteins
;
Protein separation
;
Protein sequencing
Source:
Springer Online Journal Archives 1860-2000
Topics:
Chemistry and Pharmacology
Notes:
Summary Since ribosomes are a fundamental feature component of all organisms, they present a good model for studying evolutionary diversity. We investigated the ribosomal proteins of the archaebacteriumSulfolobus acidocaldarius, which contains slightly more ribosomal proteins thanEscherichia coli. While the ribosomal proteins of most organisms contain a high proportion of lysine and arginine residues, these basic amino acids are particularly prevelent in the thermaacidophile organismSulfolobus, a possible reason for poor separations of total ribosomal proteins ofS. acidocaldarius by single column HPLC. To solve this complex separation problem, we developed the discontinuous reversed-phase HPLC (Disc RPC) method. Discontinuus chromatography combines at least two different stationary phases in a sequence of increasing retention times for the elution of proteins. Unlike other multi-column techniques, all of the injected sample passes through this discontinous stationary phase, which is used in place of single columns, thus permitting separations to be carried out without the need any changes in the HPLC system. Here we present Disc RPC separations of 30S ribosomal proteins fromS. acidocaldarius with a variety of stationary phases of different legnths and diameters. Each of five Disc RPC categories presented in this paper has some special application possibilities when used in protein HPLC.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF02316443
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