Abstract：
:We have recently identified a new family of multidomain oxidoreductase (redox) enzymes, the MICALs, that directly regulate the actin cytoskeletal elements necessary for the morphology, motility, and trajectory of cells. Our genetic assays reveal that Mical is both necessary and sufficient for actin organization and cellular effects in vivo and our biochemical assays with purified Mical protein reveal that Mical utilizes its redox activity to directly disassemble actin filaments. These results identify Mical proteins as novel actin disassembly factors and uncover a redox signaling mechanism that directly regulates the actin cytoskeleton. These results have also set the stage for in-depth characterization of the Mical enzyme. However, it has been difficult to obtain sufficient amounts of highly-pure Mical protein to conduct further biochemical, structural, imaging, catalytic, and other high-precision studies. Herein, we describe a means for expressing high levels of soluble recombinant Mical protein in bacteria. Likewise, we have designed a new purification strategy that enables the rapid and efficient purification of milligram quantities of highly-pure and >99% active Mical protein. This new strategy for generating large amounts of highly-pure and active Mical protein will aid research objectives designed to characterize the biochemical, enzymology, and structural biology of Mical and its effects on actin filament dynamics.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Wu H,Hung RJ,Terman JRdoi
10.1016/j.pep.2016.05.008subject
Has Abstractpub_date
2016-11-01 00:00:00pages
116-124eissn
1046-5928issn
1096-0279pii
S1046-5928(16)30086-9journal_volume
127pub_type
杂志文章abstract：:The bivalent anti-human anti-T cell immunotoxin A-dmDT390-bisFv(G(4)S) was developed for treatment of T cell leukemia, autoimmune diseases, and tolerance induction for transplantation. The multi-domain structure of the bivalent immunotoxin hinders efficient production in Escherichia coli and most eukaryotes are sensit...
journal_title：Protein expression and purification
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abstract：:High-level constitutive expression of xylanase GH11 from Penicillium occitanis Pol6 termed PoXyn2 was achieved using the methylotrophic yeast Pichia pastoris. The PoXyn2 cDNA encoding for a mature xylanase of 320 amino acids was subcloned into the pGAPZαA vector, to construct recombinant xylanse with six histidine res...
journal_title：Protein expression and purification
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journal_title：Protein expression and purification
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1996.0047
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journal_title：Protein expression and purification
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journal_title：Protein expression and purification
pub_type： 杂志文章
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journal_title：Protein expression and purification
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doi：10.1016/j.pep.2014.07.003
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1995.1070
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journal_title：Protein expression and purification
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journal_title：Protein expression and purification
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doi：10.1016/j.pep.2014.09.015
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2004.10.003
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2006.10.005
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1997.0757
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2014.03.011
更新日期：2014-07-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.2001.1470
更新日期：2001-10-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.2001.1402
更新日期：2001-04-01 00:00:00
abstract：:A tremendous increase has taken place over the last decades in the biochemical and clinical use of antibodies. Unfortunately, the constantly growing demand has not been matched by a corresponding easy access to pure immunoglobulin, as purification procedures tend to be either laborious, expensive, or inefficient. We p...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/s1046-5928(03)00130-x
更新日期：2003-08-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/s1046-5928(02)00019-0
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2007.03.016
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journal_title：Protein expression and purification
pub_type： 杂志文章
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journal_title：Protein expression and purification
pub_type： 杂志文章
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journal_title：Protein expression and purification
pub_type： 杂志文章
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2004.12.003
更新日期：2005-04-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2020.105649
更新日期：2020-09-01 00:00:00
abstract：:Telomerase is a specialized reverse transcriptase that catalyzes the addition of telomeric repeats, TTAGGG in all vertebrates, to the ends of chromosomes. The lack of recombinant purified human telomerase reverse transcriptase (hTERT) has hampered biochemical and structural studies. The primary problem in generating a...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2007.05.016
更新日期：2007-11-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1998.0969
更新日期：1998-12-01 00:00:00
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journal_title：Protein expression and purification
pub_type： 杂志文章
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abstract：:Fab fragments isolated from papain digests of monoclonal antibodies have a wide variety of uses in analytical and in both in vivo and in vitro diagnostic applications. A novel, non-affinity method which uses the Gradiflow to purify Fab fragments from the papain digest of a mouse IgG1 anti-c-myc monoclonal antibody is ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2003.07.005
更新日期：2003-12-01 00:00:00
abstract：:The YUH1 gene coding for ubiquitin C-terminal hydrolase 1, a deubiquitinating enzyme, was cloned from the Saccharomyces cerevisiae genomic DNA and expressed in Escherichia coli. YUH1 was fused with the 6 histidine tag at the N-terminus (H6YUH1) or C-terminus (YUH1H6) and purified by an immobilized metal affinity chrom...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2007.07.005
更新日期：2007-11-01 00:00:00
