Scopus Publication Detail
The publication detail shows the title, authors (with indicators showing other profiled authors), information on the publishing organization, abstract and a link to the article in Scopus. This abstract is what is used to create the fingerprint of the publication.
Expression and purification of recombinant Saccharomyces cerevisiae mitochondrial carrier protein YGR257Cp (Mtm1p)
Mei M. Whittaker; James W. Whittaker(Profiled Author: James Whittaker)
Protein Expression and Purification. 2014;93:77-86.Abstract
The Saccharomyces cerevisiae mitochondrial carrier YGR257Cp (Mtm1p) is an integral membrane protein that plays an essential role in mitochondrial iron homeostasis and respiratory functions, but its carrier substrate has not previously been identified. Large amounts of pure protein are required for biochemical characterization, including substrate screening. Functional complementation of a Saccharomyces knockout by expression of TwinStrep tagged YGR257Cp demonstrates that an affinity tag does not interfere with protein function, but the expression level is very low. Heterologous expression in Pichia pastoris improves the yield but the product is heterogeneous. Expression has been screened in several Escherichia coli hosts, optimizing yield by modifying induction conditions and supplementing with rare tRNAs to overcome codon bias in the eukaryotic gene. Detection of an additional N-terminal truncation product in E. coli reveals the presence of a secondary intracistronic translation initiation site, which can be eliminated by silent mutagenesis of an alternative (Leu) initiation codon, resulting in production of a single, full-length polypeptide (∼30% of the total protein) as insoluble inclusion bodies. Purified inclusion bodies were successfully refolded and affinity purified, yielding approximately 40 mg of pure, soluble product per liter of culture. Refolded YGR257Cp binds pyridoxal 5′-phosphate tightly (K
This section shows information related to the publication - computed using the fingerprint of the publication - including related publications, related experts with fingerprints representing significant amounts of overlap between their fingerprint and this publication. The red dots indicate whether those experts or terms appear within the publication, thereby showing potential and actual connections.
Maja Chloupková; Linda S. LeBard; David M. KoellerJournal of Molecular Biology. 2003;331(1):155-165.
Hoon Ryu; Junghee Lee; Soren Impey; Rajiv R. Ratan; Robert J. FerranteProceedings of the National Academy of Sciences of the United States of America. 2005;102(39):13915-13920.
Xiao Qiu Xiao; Kevin L. Grove; Bernadette E. Grayson; M. Susan SmithEndocrinology. 2004;145(2):830-838.
Appears in this Document