The Nano-tag, a streptavidin-binding peptide for the purification and detection of recombinant proteins

2004 ◽  
Vol 33 (1) ◽  
pp. 39-47 ◽  
Author(s):  
Thorsten Lamla ◽  
Volker A Erdmann
Keyword(s):  
Recombinant Proteins ◽  
Binding Peptide ◽  
Streptavidin Binding

One-Step Purification of Recombinant Proteins Using a Nanomolar-Affinity Streptavidin-Binding Peptide, the SBP-Tag

2001 ◽  
Vol 23 (3) ◽  
pp. 440-446 ◽  
Author(s):  
Anthony D. Keefe ◽  
David S. Wilson ◽  
Burckhard Seelig ◽  
Jack W. Szostak
Keyword(s):  
Recombinant Proteins ◽  
Binding Peptide ◽  
One Step ◽  
Streptavidin Binding

scholarly journals Development of a nanobody tagged with streptavidin-binding peptide and its application in a Luminex fluoroimmunoassay for alpha fetal protein in serum

RSC Advances ◽  
2020 ◽  
Vol 10 (40) ◽  
pp. 23767-23774
Author(s):  
Qi Chen ◽  
Danyang Sun ◽  
Hua Pei ◽  
Benchao Su ◽  
Kunlu Bao ◽  
...  
Keyword(s):  
Fusion Protein ◽  
Binding Peptide ◽  
Immunological Diagnosis ◽  
Streptavidin Binding

A nanobody/streptavidin-binding peptide fusion protein was developed and proved to be a very promising immunological diagnosis reagent for disease-related biomarkers.

scholarly journals Control of protein trafficking by reversible masking of transport signals

2016 ◽  
Vol 27 (8) ◽  
pp. 1310-1319 ◽  
Author(s):  
Omer Abraham ◽  
Karnit Gotliv ◽  
Anna Parnis ◽  
Gaelle Boncompain ◽  
Franck Perez ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Protein Trafficking ◽  
Protein Transport ◽  
Protein Import ◽  
Binding Peptide ◽  
Protein Traffic ◽  
Alternative Approach ◽  
Targeting Signal ◽  
Regulated Trafficking ◽  
Streptavidin Binding

Systems that allow the control of protein traffic between subcellular compartments have been valuable in elucidating trafficking mechanisms. Most current approaches rely on ligand or light-controlled dimerization, which results in either retardation or enhancement of the transport of a reporter. We developed an alternative approach for trafficking regulation that we term “controlled unmasking of targeting elements” (CUTE). Regulated trafficking is achieved by reversible masking of the signal that directs the reporter to its target organelle, relying on the streptavidin–biotin system. The targeting signal is generated within or immediately after a 38–amino acid streptavidin-binding peptide (SBP) that is appended to the reporter. The binding of coexpressed streptavidin to SBP causes signal masking, whereas addition of biotin causes complex dissociation and triggers protein transport to the target organelle. We demonstrate the application of this approach to the control of nuclear and peroxisomal protein import and the generation of biotin-dependent trafficking through the endocytic and COPI systems. By simultaneous masking of COPI and endocytic signals, we were able to generate a synthetic pathway for efficient transport of a reporter from the plasma membrane to the endoplasmic reticulum.

scholarly journals Heparin-binding peptide as a novel affinity tag for purification of recombinant proteins

2016 ◽  
Vol 126 ◽  
pp. 93-103 ◽  
Author(s):  
Jacqueline Morris ◽  
Srinivas Jayanthi ◽  
Rebekah Langston ◽  
Anna Daily ◽  
Alicia Kight ◽  
...  
Keyword(s):  
Recombinant Proteins ◽  
Heparin Binding ◽  
Affinity Tag ◽  
Binding Peptide

Controlled presentation of recombinant proteins via a zinc-binding peptide-linker in two and three dimensional formats

Biomaterials ◽  
2009 ◽  
Vol 30 (34) ◽  
pp. 6614-6620 ◽  
Author(s):  
Michael R. Doran ◽  
Brandon D. Markway ◽  
Tristan I. Croll ◽  
Sergio Sara ◽  
Trent P. Munro ◽  
...  
Keyword(s):  
Recombinant Proteins ◽  
Three Dimensional ◽  
Zinc Binding ◽  
Binding Peptide

scholarly journals Membrane translocation of lumenal domains of membrane proteins powered by downstream transmembrane sequences

2013 ◽  
Vol 24 (19) ◽  
pp. 3123-3132 ◽  
Author(s):  
Takaaki Yabuki ◽  
Fumiko Morimoto ◽  
Yuichiro Kida ◽  
Masao Sakaguchi
Keyword(s):  
Endoplasmic Reticulum ◽  
Membrane Proteins ◽  
Endoplasmic Reticulum Membrane ◽  
Type I ◽  
Surface Plasmon Resonance Analysis ◽  
Binding Peptide ◽  
Resonance Analysis ◽  
N Terminus ◽  
Signal Anchor ◽  
Streptavidin Binding

Translocation of the N-terminus of a type I signal anchor (SA-I) sequence across the endoplasmic reticulum membrane can be arrested by tagging with a streptavidin-binding peptide tag (SBP tag) and trapping by streptavidin. In the present study, we first examine the affinity required for the translocation arrest. When the SBP tag is serially truncated, the ability for arrest gradually decreases. Surface plasmon resonance analysis shows that an interaction as strong as 10−8 M or a smaller dissociation constant is required for trapping the topogenesis of a natural SA-I sequence. Such truncated tags, however, become effective by mutating the SA-I sequence, suggesting that the translocation motivation is considerably influenced by the properties of the SA-I sequence. In addition, we introduce the SBP tag into lumenal loops of a multispanning membrane protein, human erythrocyte band 3. Among the tagged loops between transmembrane 1 (TM1) and TM8, three loops are trapped by cytosolic streptavidin. These loops are followed by TM sequences possessing topogenic properties, like the SA-I sequence, and translocation of one loop is diminished by insertion of a proline into the following TM sequence. These findings suggest that the translocation of lumenal loops by SA-I–like TM sequences has a crucial role in topogenesis of multispanning membrane proteins.

scholarly journals Two translocating hydrophilic segments of a nascent chain span the ER membrane during multispanning protein topogenesis

2007 ◽  
Vol 179 (7) ◽  
pp. 1441-1452 ◽  
Author(s):  
Yuichiro Kida ◽  
Fumiko Morimoto ◽  
Masao Sakaguchi
Keyword(s):  
Type I ◽  
Free System ◽  
Binding Peptide ◽  
Terminal Domain ◽  
N Terminus ◽  
Nascent Chain ◽  
A Cell ◽  
Signal Anchor ◽  
Anchor Sequence ◽  
Streptavidin Binding

During protein integration into the endoplasmic reticulum, the N-terminal domain preceding the type I signal-anchor sequence is translocated through a translocon. By fusing a streptavidin-binding peptide tag to the N terminus, we created integration intermediates of multispanning membrane proteins. In a cell-free system, N-terminal domain (N-domain) translocation was arrested by streptavidin and resumed by biotin. Even when N-domain translocation was arrested, the second hydrophobic segment mediated translocation of the downstream hydrophilic segment. In one of the defined intermediates, two hydrophilic segments and two hydrophobic segments formed a transmembrane disposition in a productive state. Both of the translocating hydrophilic segments were crosslinked with a translocon subunit, Sec61α. We conclude that two translocating hydrophilic segment in a single membrane protein can span the membrane during multispanning topogenesis flanking the translocon. Furthermore, even after six successive hydrophobic segments entered the translocon, N-domain translocation could be induced to restart from an arrested state. These observations indicate the remarkably flexible nature of the translocon.

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