Folding in vitro of bovine pancreatic trypsin inhibitor in the presence of proteins of the endoplasmic reticulum

1992 ◽  
Vol 14 (1) ◽  
pp. 10-15 ◽  
Author(s):  
André Zapun ◽  
Thomas E. Creighton ◽  
Pamela J. E. Rowling ◽  
Robert B. Freedman
Keyword(s):  
Endoplasmic Reticulum ◽  
Trypsin Inhibitor ◽  
Bovine Pancreatic Trypsin Inhibitor ◽  
Pancreatic Trypsin Inhibitor

Conformational changes of the chaperone SecB upon binding to a model substrate – bovine pancreatic trypsin inhibitor (BPTI)

2011 ◽  
Vol 392 (10) ◽  
pp. 849-858 ◽  
Author(s):  
Michaela M. Haimann ◽  
Yasar Akdogan ◽  
Reinhard Philipp ◽  
Raghavan Varadarajan ◽  
Dariush Hinderberger ◽  
...  
Keyword(s):  
Conformational Changes ◽  
Trypsin Inhibitor ◽  
Tertiary Structure ◽  
Substrate Binding ◽  
Spin Labeling ◽  
Bovine Pancreatic Trypsin Inhibitor ◽  
Spin Labels ◽  
Distance Measurements ◽  
Pancreatic Trypsin Inhibitor

Abstract SecB is a homotetrameric cytosolic chaperone that forms part of the protein translocation machinery in E. coli. Due to SecB, nascent polypeptides are maintained in an unfolded translocation-competent state devoid of tertiary structure and thus are guided to the translocon. In vitro SecB rapidly binds to a variety of ligands in a non-native state. We have previously investigated the bound state conformation of the model substrate bovine pancreatic trypsin inhibitor (BPTI) as well as the conformation of SecB itself by using proximity relationships based on site-directed spin labeling and pyrene fluorescence methods. It was shown that SecB undergoes a conformational change during the process of substrate binding. Here, we generated SecB mutants containing but a single cysteine per subunit or an exposed highly reactive new cysteine after removal of the nearby intrinsic cysteines. Quantitative spin labeling was achieved with the methanethiosulfonate spin label (MTS) at positions C97 or E90C, respectively. Highfield (W-band) electron paramagnetic resonance (EPR) measurements revealed that with BPTI present the spin labels are exposed to a more polar/hydrophilic environment. Nanoscale distance measurements with double electron-electron resonance (DEER) were in excellent agreement with distances obtained by molecular modeling. Binding of BPTI also led to a slight change in distances between labels at C97 but not at E90C. While the shorter distance in the tetramer increased, the larger diagonal distance decreased. These findings can be explained by a widening of the tetrameric structure upon substrate binding much like the opening of two pairs of scissors.

scholarly journals Screening for Stable Mutants with Amino Acid Pairs Substituted for the Disulfide Bond between Residues 14 and 38 of Bovine Pancreatic Trypsin Inhibitor (BPTI)

2002 ◽  
Vol 277 (52) ◽  
pp. 51043-51048 ◽  
Author(s):  
Yoshihisa Hagihara ◽  
Kentaro Shiraki ◽  
Tsutomu Nakamura ◽  
Koichi Uegaki ◽  
Masahiro Takagi ◽  
...  
Keyword(s):  
Amino Acid ◽  
Disulfide Bond ◽  
Trypsin Inhibitor ◽  
Bovine Pancreatic Trypsin Inhibitor ◽  
Pancreatic Trypsin Inhibitor

Synthetic Approaches to Disulfide-free Circular Bovine Pancreatic Trypsin Inhibitor (c-BPTI) Analogues

2006 ◽  
Vol 12 (2) ◽  
pp. 93-104 ◽  
Author(s):  
Judit Tulla-Puche ◽  
Irina V. Getun ◽  
Yvonne M. Angell ◽  
Jordi Alsina ◽  
Fernando Albericio ◽  
...  
Keyword(s):  
Trypsin Inhibitor ◽  
Bovine Pancreatic Trypsin Inhibitor ◽  
Pancreatic Trypsin Inhibitor ◽  
Synthetic Approaches

A Pulsed-Field Gradient NMR Study of Bovine Pancreatic Trypsin Inhibitor Self-Association†

Biochemistry ◽  
1997 ◽  
Vol 36 (11) ◽  
pp. 3383-3388 ◽  
Author(s):  
Elena Ilyina ◽  
Vikram Roongta ◽  
Hong Pan ◽  
Clare Woodward ◽  
Kevin H. Mayo
Keyword(s):  
Field Gradient ◽  
Trypsin Inhibitor ◽  
Bovine Pancreatic Trypsin Inhibitor ◽  
Pulsed Field Gradient Nmr ◽  
Pulsed Field ◽  
Nmr Study ◽  
Pancreatic Trypsin Inhibitor ◽  
Self Association
Sign in / Sign up

Export Citation Format

Share Document