scholarly journals Production of dye-binding esterase reactor after separation and detection using combined native isoelectric focusing and blue native electrophoresis

2020 ◽  
Vol 64 (1) ◽  
pp. 1-6
Author(s):  
Youji Shimazaki ◽  
Hikari Nakamaru
Keyword(s):  
Isoelectric Focusing ◽  
Dye Binding ◽  
Native Electrophoresis ◽  
Blue Native Electrophoresis ◽  
Blue Native

scholarly journals Tim18p Is a New Component of the Tim54p-Tim22p Translocon in the Mitochondrial Inner Membrane

2000 ◽  
Vol 11 (1) ◽  
pp. 103-116 ◽  
Author(s):  
Oliver Kerscher ◽  
Naresh B. Sepuri ◽  
Robert E. Jensen
Keyword(s):  
Growth Defect ◽  
Inner Membrane ◽  
New Members ◽  
Mitochondrial Inner Membrane ◽  
Native Electrophoresis ◽  
Multiple Copies ◽  
New Gene ◽  
Synthetically Lethal ◽  
Blue Native Electrophoresis ◽  
Blue Native

The mitochondrial inner membrane contains two separate translocons: one required for the translocation of matrix-targeted proteins (the Tim23p-Tim17p complex) and one for the insertion of polytopic proteins into the mitochondrial inner membrane (the Tim54p-Tim22p complex). To identify new members of the Tim54p-Tim22p complex, we screened for high-copy suppressors of the temperature-sensitivetim54-1 mutant. We identified a new gene,TIM18, that encodes an integral protein of the inner membrane. The following genetic and biochemical observations suggest that the Tim18 protein is part of the Tim54p-Tim22p complex in the inner membrane: multiple copies of TIM18 suppress thetim54-1 growth defect; thetim18::HIS3 disruption is synthetically lethal with tim54-1; Tim54p and Tim22p can be coimmune precipitated with the Tim18 protein; and Tim18p, along with Tim54p and Tim22p, is detected in an ∼300-kDa complex after blue native electrophoresis. We propose that Tim18p is a new component of the Tim54p-Tim22p machinery that facilitates insertion of polytopic proteins into the mitochondrial inner membrane.

High throughput two-dimensional blue-native electrophoresis: A tool for functional proteomics of mitochondria and signaling complexes

PROTEOMICS ◽  
2002 ◽  
Vol 2 (8) ◽  
pp. 969 ◽  
Author(s):  
Paul S. Brookes ◽  
Anita Pinner ◽  
Anup Ramachandran ◽  
Lori Coward ◽  
Stephen Barnes ◽  
...  
Keyword(s):  
High Throughput ◽  
Functional Proteomics ◽  
Two Dimensional ◽  
Native Electrophoresis ◽  
Blue Native Electrophoresis ◽  
Blue Native

Blue native electrophoresis as a tool for detection of PIP-containing protein complexes in the plasma membranes

2013 ◽  
Vol 60 (4) ◽  
pp. 563-570
Author(s):  
T. A. Shevyreva ◽  
M. S. Piotrovskii ◽  
B. V. Belugin ◽  
I. M. Zhestkova ◽  
M. S. Trofimova
Keyword(s):  
Plasma Membranes ◽  
Protein Complexes ◽  
Native Electrophoresis ◽  
Blue Native Electrophoresis ◽  
Blue Native

scholarly journals Mitochondrial Import of the ADP/ATP Carrier: the Essential TIM Complex of the Intermembrane Space Is Required for Precursor Release from the TOM Complex

2002 ◽  
Vol 22 (22) ◽  
pp. 7780-7789 ◽  
Author(s):  
Kaye N. Truscott ◽  
Nils Wiedemann ◽  
Peter Rehling ◽  
Hanne Müller ◽  
Chris Meisinger ◽  
...  
Keyword(s):  
Outer Membrane ◽  
Intermembrane Space ◽  
Hydrophobic Membrane ◽  
Tom Complex ◽  
Native Electrophoresis ◽  
Mitochondrial Intermembrane Space ◽  
First Time ◽  
Blue Native Electrophoresis ◽  
Step Mechanism ◽  
Blue Native

ABSTRACT The mitochondrial intermembrane space contains a protein complex essential for cell viability, the Tim9-Tim10 complex. This complex is required for the import of hydrophobic membrane proteins, such as the ADP/ATP carrier (AAC), into the inner membrane. Different views exist about the role played by the Tim9-Tim10 complex in translocation of the AAC precursor across the outer membrane. For this report we have generated a new tim10 yeast mutant that leads to a strong defect in AAC import into mitochondria. Thereby, for the first time, authentic AAC is stably arrested in the translocase complex of the outer membrane (TOM), as shown by antibody shift blue native electrophoresis. Surprisingly, AAC is still associated with the receptors Tom70 and Tom20 when the function of Tim10 is impaired. The nonessential Tim8-Tim13 complex of the intermembrane space is not involved in the transfer of AAC across the outer membrane. These results define a two-step mechanism for translocation of AAC across the outer membrane. The initial insertion of AAC into the import channel is independent of the function of Tim9-Tim10; however, completion of translocation across the outer membrane, including release from the TOM complex, requires a functional Tim9-Tim10 complex.

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