scholarly journals Availability of Dihydrofolate Reductase Affinity Handle in Expressing Human Prolactin as a Soluble Fusion Protein

1993 ◽  
Vol 57 (11) ◽  
pp. 1955-1957 ◽  
Author(s):  
Masahiro Iwakura ◽  
Minoru Morikawa
Keyword(s):  
Fusion Protein ◽  
Dihydrofolate Reductase ◽  
Human Prolactin ◽  
Soluble Fusion Protein

Aggregation kinetics in the presence of brain lipids of Aβ(1–40) cleaved from a soluble fusion protein

2018 ◽  
Vol 1860 (9) ◽  
pp. 1681-1686 ◽  
Author(s):  
Miriam A. Kael ◽  
Daniel K. Weber ◽  
Frances Separovic ◽  
Marc-Antoine Sani
Keyword(s):  
Fusion Protein ◽  
Aggregation Kinetics ◽  
Brain Lipids ◽  
Soluble Fusion Protein

Overexpression and Large-Scale Purification of Recombinant Hamster Polymorphic ArylamineN-Acetyltransferase as a Dihydrofolate Reductase Fusion Protein

1997 ◽  
Vol 10 (1) ◽  
pp. 141-153 ◽  
Author(s):  
Kristina R.K. Sticha ◽  
Christine A. Sieg ◽  
Carl P. Bergstrom ◽  
Patrick E. Hanna ◽  
Carston R. Wagner
Keyword(s):  
Fusion Protein ◽  
Dihydrofolate Reductase ◽  
Large Scale

scholarly journals Recombinant Adenovirus Expressing a Soluble Fusion Protein PD-1/CD137L Subverts the Suppression of CD8+ T Cells in HCC

2019 ◽  
Vol 27 (11) ◽  
pp. 1906-1918 ◽  
Author(s):  
Yonghui Zhang ◽  
Hailin Zhang ◽  
Mei Wei ◽  
Tao Mou ◽  
Tao Shi ◽  
...  
Keyword(s):  
T Cells ◽  
Fusion Protein ◽  
Cd8 T Cells ◽  
Recombinant Adenovirus ◽  
Soluble Fusion Protein

scholarly journals Import of a DHFR hybrid protein into glycosomes in vivo is not inhibited by the folate-analogue aminopterin.

1996 ◽  
Vol 132 (3) ◽  
pp. 311-324 ◽  
Author(s):  
T Häusler ◽  
Y D Stierhof ◽  
E Wirtz ◽  
C Clayton
Keyword(s):  
Fusion Protein ◽  
Trypanosoma Brucei ◽  
Dihydrofolate Reductase ◽  
Fusion Proteins ◽  
Phosphoglycerate Kinase ◽  
Cytochrome Oxidase Subunit ◽  
Conformational Properties ◽  
Terminal Amino ◽  
Carboxy Terminal

Dihydrofolate reductase fusion proteins have been widely used to study conformational properties of polypeptides translocated across membranes. We have studied the import of dihydrofolate reductase fusion proteins into glycosomes and mitochondria of Trypanosoma brucei. As signal sequences we used the last 22 carboxy-terminal amino acids of glycosomal phosphoglycerate kinase for glycosomes, and the cleavable presequences of yeast cytochrome b2 or cytochrome oxidase subunit IV for mitochondria. Upon addition of aminopterin, a folate analogue that stabilizes the dihydrofolate reductase moiety, import of the fusion protein targeted to glycosomes was not inhibited, although the results of protease protection assays showed that the fusion protein could bind the drug. Under the same conditions, import of a DHFR fusion protein targeted to mitochondria was inhibited by aminopterin. When DHFR fusion proteins targeted simultaneously to both glycosomes and mitochondria were expressed, import into mitochondria was inhibited by aminopterin, whereas uptake of the same proteins into glycosomes was either unaffected or slightly increased. These findings suggest that the glycosomes possess either a strong unfolding activity or an unusually large or flexible translocation channel.

scholarly journals Ability of methotrexate to inhibit translocation to the cytosol of dihydrofolate reductase fused to diphtheria toxin

1996 ◽  
Vol 313 (2) ◽  
pp. 647-653 ◽  
Author(s):  
Olav KLINGENBERG ◽  
Sjur OLSNES
Keyword(s):  
Fusion Protein ◽  
Dihydrofolate Reductase ◽  
Diphtheria Toxin ◽  
In Vitro Transcription ◽  
Wild Type ◽  
Toxin B ◽  
Rabbit Reticulocyte Lysate System ◽  
Reticulocyte Lysate ◽  
Diphtheria Toxin A

A fusion protein consisting of dihydrofolate reductase and diphtheria toxin A-fragment was made by genetically linking cDNA for the two proteins followed by in vitro transcription and translation in a rabbit reticulocyte lysate system. The dihydrofolate reductase in the fusion protein exhibited enzyme activity and, in the presence of methotrexate which imposes a tight structure on dihydrofolate reductase, it was trypsin resistant, indicating that it was correctly folded. When reconstituted with diphtheria toxin B-fragment, it bound specifically to diphtheria toxin receptors and was translocated into cells upon exposure to low pH. Methotrexate prevented the translocation. Protein synthesis was inhibited in cells incubated with the reconstituted fusion protein, but the inhibition was reduced in the presence of methotrexate. We also made a fusion protein containing a mutated dihydrofolate reductase with much lower affinity to methotrexate. Methotrexate did not prevent translocation of this protein. The data indicate that methotrexate prevents translocation of the fusion protein containing wild-type dihydrofolate reductase by imposing a tight structure on to the enzyme.

scholarly journals Resistance to Methotrexate Due to AcrAB-Dependent Export from Escherichia coli

2000 ◽  
Vol 44 (11) ◽  
pp. 3210-3212 ◽  
Author(s):  
Stephan J. Kopytek ◽  
John C. D. Dyer ◽  
Gwendowlyn S. Knapp ◽  
James C. Hu
Keyword(s):  
Escherichia Coli ◽  
Multidrug Resistance ◽  
Fusion Protein ◽  
Dihydrofolate Reductase ◽  
Efflux Pump ◽  
In Cells

ABSTRACT Many laboratory strains of Escherichia coli are resistant to methotrexate (MTX), a folate analogue that binds dihydrofolate reductase (DHFR). Mutations that inactivate eithertolC or acrA confer MTX sensitivity. Further, overexpression of a fusion protein with DHFR activity reverses this sensitivity by titrating out intracellular MTX. These results suggest that MTX accumulates in cells where mutations inacrA or tolC have inactivated the TolC-dependent AcrAB multidrug resistance efflux pump.

scholarly journals Human Metapneumovirus Reinfection among Children in Thailand Determined by ELISA Using Purified Soluble Fusion Protein

2008 ◽  
Vol 198 (6) ◽  
pp. 836-842 ◽  
Author(s):  
Julie A. Pavlin ◽  
Andrew C. Hickey ◽  
Nancy Ulbrandt ◽  
Yee‐Peng Chan ◽  
Timothy P. Endy ◽  
...  
Keyword(s):  
Fusion Protein ◽  
Human Metapneumovirus ◽  
Soluble Fusion Protein
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