Disulfide bond contribution to protein stability: Positional effects of substitution in the hydrophobic core of the two-stranded .alpha.-helical coiled-coil

Biochemistry ◽  
1993 ◽  
Vol 32 (12) ◽  
pp. 3178-3187 ◽  
Author(s):  
Nian E. Zhou ◽  
Cyril M. Kay ◽  
Robert S. Hodges
Keyword(s):  
Protein Stability ◽  
Disulfide Bond ◽  
Coiled Coil ◽  
Hydrophobic Core ◽  
Positional Effects

scholarly journals A comprehensive analysis of novel disulfide bond introduction site into the constant domain of human Fab

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Hitomi Nakamura ◽  
Moeka Yoshikawa ◽  
Naoko Oda-Ueda ◽  
Tadashi Ueda ◽  
Takatoshi Ohkuri
Keyword(s):  
Thermal Stability ◽  
Pichia Pastoris ◽  
Protein Stability ◽  
Disulfide Bond ◽  
Binding Activity ◽  
Comprehensive Analysis ◽  
The Other ◽  
Constant Domain ◽  
Intermolecular Disulfide Bond ◽  
Sds Page

AbstractGenerally, intermolecular disulfide bond contribute to the conformational protein stability. To identify sites where intermolecular disulfide bond can be introduced into the Fab’s constant domain of the therapeutic IgG, Fab mutants were predicted using the MOE software, a molecular simulator, and expressed in Pichia pastoris. SDS-PAGE analysis of the prepared Fab mutants from P. pastoris indicated that among the nine analyzed Fab mutants, the F130C(H):Q124C(L), F174C(H):S176C(L), V177C(H):Q160C(L), F174C(H):S162C(L), F130C(H):S121C(L), and A145C(H):F116C(L) mutants mostly formed intermolecular disulfide bond. All these mutants showed increased thermal stability compared to that of Fab without intermolecular disulfide bond. In the other mutants, the intermolecular disulfide bond could not be completely formed, and the L132C(H):F118C(L) mutant showed only a slight decrease in binding activity and β-helix content, owing to the exertion of adverse intermolecular disulfide bond effects. Thus, our comprehensive analysis reveals that the introduction of intermolecular disulfide bond in the Fab’s constant domain is possible at various locations. These findings provide important insights for accomplishing human Fab stabilization.

scholarly journals 2P-079 Influences of the disulfide bond of the starch-binding domain of glucoamylase on the protein stability and refolding(The 46th Annual Meeting of the Biophysical Society of Japan)

2008 ◽  
Vol 48 (supplement) ◽  
pp. S87
Author(s):  
Hayuki Sugimoto ◽  
Miho Nakaura ◽  
Shigenori Nishimura ◽  
Shuichi Karita ◽  
Hideo Miyake ◽  
...  
Keyword(s):  
Protein Stability ◽  
Annual Meeting ◽  
Disulfide Bond ◽  
Binding Domain ◽  
Biophysical Society

scholarly journals In VivoImpact of Met221 Substitution in GOB Metallo-β-Lactamase

2012 ◽  
Vol 56 (4) ◽  
pp. 1769-1773 ◽  
Author(s):  
Jorgelina Morán-Barrio ◽  
María-Natalia Lisa ◽  
Alejandro J. Vila
Keyword(s):  
Antibiotic Resistance ◽  
Protein Stability ◽  
Metal Binding ◽  
Active Sites ◽  
Bacterial Resistance ◽  
Structural Diversity ◽  
Hydrophobic Core

ABSTRACTMetallo-β-lactamases (MβLs) represent one of the main mechanisms of bacterial resistance against β-lactam antibiotics. The elucidation of their mechanism has been limited mostly by the structural diversity among their active sites. All MβLs structurally characterized so far present a Cys or a Ser residue at position 221, which is critical for catalysis. GOB lactamases stand as an exception within this picture, possessing a Met residue in this location. We studied different mutants in this position, and we show that Met221 is essential for protein stability, most likely due to its involvement in a hydrophobic core. In contrast to other known MβLs, residue 221 is not involved in metal binding or in catalysis in GOB enzymes, further highlighting the structural diversity of MβLs. We also demonstrate the usefulness of protein periplasmic profiles to assess the contribution of protein stability to antibiotic resistance.

scholarly journals Effects of an Interchain Disulfide Bond on Tropomyosin Structure: A Molecular Dynamics Study

2018 ◽  
Vol 19 (11) ◽  
pp. 3376 ◽  
Author(s):  
Natalia A. Koubassova ◽  
Sergey Y. Bershitsky ◽  
Andrey K. Tsaturyan
Keyword(s):  
Heart Failure ◽  
Molecular Dynamics ◽  
Disulfide Bond ◽  
Coiled Coil ◽  
Middle Part ◽  
Actin Binding ◽  
Cross Linking ◽  
Interchain Disulfide Bond ◽  
Structural And Functional Properties ◽  
The Cross

Tropomyosin (Tpm) is a coiled-coil actin-binding dimer protein that participates in the regulation of muscle contraction. Both Tpm chains contain Cys190 residues which are normally in the reduced state, but form an interchain disulfide bond in failing heart. Changes in structural and functional properties of Tpm and its complexes with actin upon disulfide cross-linking were studied using various experimental methods. To understand the molecular mechanism underlying these changes and to reveal the possible mechanism of the involvement of the cross-linking in heart failure, molecular dynamics (MD) simulations of the middle part of Tpm were performed in cross-linked and reduced states. The cross-linking increased bending stiffness of Tpm assessed from MD trajectories at 27 °C in agreement with previous experimental observations. However, at 40 °C, the cross-linking caused a decrease in Tpm stiffness and a significant reduction in the number of main chain hydrogen bonds in the vicinity of residues 133 and 134. These data are in line with observations showing enhanced thermal unfolding of the least stable part of Tpm at 30–40 °C and accelerated trypsin cleavage at residue 133 at 40 °C (but not at 27 °C) upon cross-linking. These results allow us to speculate about the possible mechanism of involvement of Tpm cross-linking to heart failure pathogenesis.

Retention of Coiled-Coil Dimer Formation in the Absence of Ion Pairing at Positions Flanking the Hydrophobic Core

Biochemistry ◽  
2019 ◽  
Vol 58 (48) ◽  
pp. 4821-4826 ◽  
Author(s):  
Naomi A. Biok ◽  
Alexander D. Passow ◽  
Chenxuan Wang ◽  
Craig A. Bingman ◽  
Nicholas L. Abbott ◽  
...  
Keyword(s):  
Coiled Coil ◽  
Ion Pairing ◽  
Hydrophobic Core ◽  
Dimer Formation

Engineering of the hydrophobic core of an α-helical coiled coil

Biopolymers ◽  
2000 ◽  
Vol 55 (5) ◽  
pp. 407-414 ◽  
Author(s):  
Tomohiro Kiyokawa ◽  
Kenji Kanaori ◽  
Kunihiko Tajima ◽  
Toshiki Tanaka
Keyword(s):  
Coiled Coil ◽  
Hydrophobic Core

scholarly journals Conserved Asp-137 Is Important for both Structure and Regulatory Functions of Cardiac α-Tropomyosin (α-TM) in a Novel Transgenic Mouse Model Expressing α-TM-D137L

2013 ◽  
Vol 288 (23) ◽  
pp. 16235-16246 ◽  
Author(s):  
Sumeyye Yar ◽  
Shamim A. K. Chowdhury ◽  
Robert T. Davis ◽  
Minae Kobayashi ◽  
Michelle M. Monasky ◽  
...  
Keyword(s):  
Mouse Model ◽  
Transgenic Mouse ◽  
Coiled Coil ◽  
Transgenic Mouse Model ◽  
Hydrophobic Core ◽  
Scanning Calorimetry ◽  
Physiological Importance ◽  
Cardiomyocyte Contractility ◽  
Regulatory Functions

α-Tropomyosin (α-TM) has a conserved, charged Asp-137 residue located in the hydrophobic core of its coiled-coil structure, which is unusual in that the residue is found at a position typically occupied by a hydrophobic residue. Asp-137 is thought to destabilize the coiled-coil and so impart structural flexibility to the molecule, which is believed to be crucial for its function in the heart. A previous in vitro study indicated that the conversion of Asp-137 to a more typical canonical Leu alters flexibility of TM and affects its in vitro regulatory functions. However, the physiological importance of the residue Asp-137 and altered TM flexibility is unknown. In this study, we further analyzed structural properties of the α-TM-D137L variant and addressed the physiological importance of TM flexibility in cardiac function in studies with a novel transgenic mouse model expressing α-TM-D137L in the heart. Our NMR spectroscopy data indicated that the presence of D137L introduced long range rearrangements in TM structure. Differential scanning calorimetry measurements demonstrated that α-TM-D137L has higher thermal stability compared with α-TM, which correlated with decreased flexibility. Hearts of transgenic mice expressing α-TM-D137L showed systolic and diastolic dysfunction with decreased myofilament Ca2+ sensitivity and cardiomyocyte contractility without changes in intracellular Ca2+ transients or post-translational modifications of major myofilament proteins. We conclude that conversion of the highly conserved Asp-137 to Leu results in loss of flexibility of TM that is important for its regulatory functions in mouse hearts. Thus, our results provide insight into the link between flexibility of TM and its function in ejecting hearts.

ph-Induced Conformational Change in An A- Helical Coiled-Coil is Controlled by His Residues in the Hydrophobic Core

2003 ◽  
Vol 10 (1) ◽  
pp. 27-33 ◽  
Author(s):  
Kiyoko Wada ◽  
Toshihisa Mizuno ◽  
Jun-ichi Oku ◽  
Toshiki Tanaka
Keyword(s):  
Conformational Change ◽  
Coiled Coil ◽  
Hydrophobic Core

Effects of single substitutions with hexafluoroleucine and trifluorovaline on the hydrophobic core formation of a heterodimeric coiled coil

2015 ◽  
Vol 175 ◽  
pp. 32-35 ◽  
Author(s):  
Susanne Huhmann ◽  
Elisabeth K. Nyakatura ◽  
Holger Erdbrink ◽  
Ulla I.M. Gerling ◽  
Constantin Czekelius ◽  
...  
Keyword(s):  
Coiled Coil ◽  
Hydrophobic Core ◽  
Core Formation

scholarly journals BPTI folding revisited: switching a disulfide into methylene thioacetal reveals a previously hidden path

2018 ◽  
Vol 9 (21) ◽  
pp. 4814-4820 ◽  
Author(s):  
Reem Mousa ◽  
Shifra Lansky ◽  
Gil Shoham ◽  
Norman Metanis
Keyword(s):  
Protein Structure ◽  
Protein Stability ◽  
Disulfide Bond ◽  
Trypsin Inhibitor ◽  
Biological Function ◽  
Bovine Pancreatic Trypsin Inhibitor ◽  
Folding Mechanism ◽  
Pancreatic Trypsin Inhibitor ◽  
Model Protein

The folding mechanism of the model protein bovine pancreatic trypsin inhibitor was revisited. By switching the solvent exposed disulfide bond with methylene thioacetal we uncovered a hidden pathway in its folding mechanism. In addition, this moiety enhanced protein stability while fully maintaining the protein structure and biological function.

Sign in / Sign up

Export Citation Format

Share Document