Phospholipase A2 Engineering. The Roles of Disulfide Bonds in Structure, Conformational Stability, and Catalytic Function

Biochemistry ◽  
1995 ◽  
Vol 34 (46) ◽  
pp. 15307-15314 ◽  
Author(s):  
Hongxin Zhu ◽  
Cynthia M. Dupureur ◽  
Xiaoyan Zhang ◽  
Ming-Daw Tsai
Keyword(s):  
Phospholipase A2 ◽  
Disulfide Bonds ◽  
Conformational Stability ◽  
Catalytic Function

Disulfide bonds of phospholipase A2 from bee venom yield discrete contributions to its conformational stability

Biochimie ◽  
2011 ◽  
Vol 93 (2) ◽  
pp. 195-201 ◽  
Author(s):  
Sylvia Welker ◽  
Yvonne Markert ◽  
Jens Köditz ◽  
Johanna Mansfeld ◽  
Renate Ulbrich-Hofmann
Keyword(s):  
Phospholipase A2 ◽  
Disulfide Bonds ◽  
Conformational Stability ◽  
Bee Venom

scholarly journals Conformational stability and activity of ribonuclease T1 with zero, one, and two intact disulfide bonds.

1988 ◽  
Vol 263 (24) ◽  
pp. 11820-11825 ◽  
Author(s):  
C N Pace ◽  
G R Grimsley ◽  
J A Thomson ◽  
B J Barnett
Keyword(s):  
Disulfide Bonds ◽  
Conformational Stability ◽  
Ribonuclease T1

scholarly journals The role of an evolutionarily conserved cis-proline in the thioredoxin-like domain of human class Alpha glutathione transferase A1-1

2003 ◽  
Vol 372 (1) ◽  
pp. 241-246 ◽  
Author(s):  
Chris NATHANIEL ◽  
Louise A. WALLACE ◽  
Jonathan BURKE ◽  
Heini W. DIRR
Keyword(s):  
Glutathione Transferase ◽  
Conformational Stability ◽  
Wild Type ◽  
Catalytic Function ◽  
Evolutionarily Conserved ◽  
Glutathione S Transferases ◽  
Equilibrium Unfolding ◽  
The Stability ◽  
Unfolding Kinetics

The thioredoxin-like fold has a βαβαββα topology, and most proteins/domains with this fold have a topologically conserved cis-proline residue at the N-terminus of β-strand 3. This residue plays an important role in the catalytic function and stability of thioredoxin-like proteins, but is reported not to contribute towards the stability of glutathione S-transferases (GSTs) [Allocati, Casalone, Masulli, Caccarelli, Carletti, Parker and Di Ilio (1999) FEBS Lett. 445, 347–350]. In order to further address the role of the cis-proline in the structure, function and stability of GSTs, cis-Pro-56 in human GST (hGST) A1-1 was replaced with a glycine, and the properties of the P56G mutant were compared with those of the wild-type protein. Not only was the catalytic function of the mutant dramatically reduced, so was its conformational stability, as indicated by equilibrium unfolding and unfolding kinetics experiments with urea as denaturant. These findings are discussed in the context of other thioredoxin-like proteins.

scholarly journals The Role of Propeptide in the Refolding of Human Group IB Phospholipase A2

2004 ◽  
Vol 36 (9) ◽  
pp. 583-588 ◽  
Author(s):  
Hong-Qiang Cheng ◽  
Gen-Jun Xu
Keyword(s):  
Light Scattering ◽  
Phospholipase A2 ◽  
Disulfide Bonds ◽  
Human Group ◽  
Terminal Sequence ◽  
E Coli ◽  
Mature Enzyme

Abstract Human group IB phospholipase A2 (IB-PLA2) and its zymogen (proIB-PLA2) were purified from E. coli. Refolding was carried out by diluting the denatured forms of both IB-PLA2 and proIB-PLA2 with renaturation buffer in which the disulfide bonds were completely reduced. The refolding yield of proIB-PLA2 was increased by about 50% over that of the mature enzyme. The refolding of IB-PLA2 usually produced aggregates under normal conditions, as determined by light scattering. In addition, the unfolding experiments showed that the mature enzyme was more stable than the proenzyme toward denaturants in the presence of DTT. Results suggested that the N-terminal sequence rather than its conformation of human proIB-PLA2 played an important role in the refolding process.

The Single Disulfide-Directed β-Hairpin Fold: Role of Disulfide Bond in Folding and Effect of an Additional Disulfide Bond on Stability

2020 ◽  
Vol 73 (4) ◽  
pp. 312
Author(s):  
Balasubramanyam Chittoor ◽  
Bankala Krishnarjuna ◽  
Rodrigo A. V. Morales ◽  
Raymond S. Norton
Keyword(s):  
Disulfide Bond ◽  
Disulfide Bonds ◽  
Solution Structure ◽  
Conformational Stability ◽  
Disulfide Bridge ◽  
Solution Nmr ◽  
Peptide Epitope ◽  
The Core ◽  
Native Fold

Disulfide bonds play a key role in the oxidative folding, conformational stability, and functional activity of many peptides. A few disulfide-rich peptides with privileged architecture such as the inhibitor cystine knot motif have garnered attention as templates in drug design. The single disulfide-directed β-hairpin (SDH), a novel fold identified more recently in contryphan-Vc1, has been shown to possess remarkable thermal, conformational, and chemical stability and can accept a short bioactive epitope without compromising the core structure of the peptide. In this study, we demonstrated that the single disulfide bond is critical in maintaining the native fold by replacing both cysteine residues with serine. We also designed an analogue with an additional, non-native disulfide bridge by replacing Gln1 and Tyr9 with Cys. Contryphan-Vc11–22[Q1C, Y9C] was synthesised utilising orthogonal cysteine protection and its solution structure determined using solution NMR spectroscopy. This analogue maintained the overall fold of native contryphan-Vc1. Previous studies had shown that the β-hairpin core of contryphan-Vc1 was resistant to proteolysis by trypsin and α-chymotrypsin but susceptible to cleavage by pepsin. Contryphan-Vc11–22[Q1C, Y9C] proved to be completely resistant to pepsin, thus confirming our design strategy. These results highlight the role of the disulfide bond in maintaining the SDH fold and provide a basis for the design of more stable analogues for peptide epitope grafting.

Comparative effect of cationic gemini surfactant and its monomeric counterpart on the conformational stability of phospholipase A2

2019 ◽  
Vol 1175 ◽  
pp. 49-55 ◽  
Author(s):  
Mehraj ud din Parray ◽  
Neha Maurya ◽  
Farooq Ahmad Wani ◽  
Mahendra S. Borse ◽  
Najmul Arfin ◽  
...  
Keyword(s):  
Phospholipase A2 ◽  
Gemini Surfactant ◽  
Conformational Stability ◽  
Comparative Effect ◽  
Cationic Gemini Surfactant

scholarly journals Human Group IIA Phospholipase A2—Three Decades on from Its Discovery

Molecules ◽  
2021 ◽  
Vol 26 (23) ◽  
pp. 7267
Author(s):  
Kieran F. Scott ◽  
Timothy J. Mann ◽  
Shadma Fatima ◽  
Mila Sajinovic ◽  
Anshuli Razdan ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Phospholipase A2 ◽  
Future Research ◽  
Human Group ◽  
Independent Function ◽  
Selective Inhibitors ◽  
Homologous Proteins ◽  
Activity Class ◽  
Catalytic Function ◽  
Catalytically Active

Phospholipase A2 (PLA2) enzymes were first recognized as an enzyme activity class in 1961. The secreted (sPLA2) enzymes were the first of the five major classes of human PLA2s to be identified and now number nine catalytically-active structurally homologous proteins. The best-studied of these, group IIA sPLA2, has a clear role in the physiological response to infection and minor injury and acts as an amplifier of pathological inflammation. The enzyme has been a target for anti-inflammatory drug development in multiple disorders where chronic inflammation is a driver of pathology since its cloning in 1989. Despite intensive effort, no clinically approved medicines targeting the enzyme activity have yet been developed. This review catalogues the major discoveries in the human group IIA sPLA2 field, focusing on features of enzyme function that may explain this lack of success and discusses future research that may assist in realizing the potential benefit of targeting this enzyme. Functionally-selective inhibitors together with isoform-selective inhibitors are necessary to limit the apparent toxicity of previous drugs. There is also a need to define the relevance of the catalytic function of hGIIA to human inflammatory pathology relative to its recently-discovered catalysis-independent function.

scholarly journals Identification of Essential Residues for the Catalytic Function of 85-kDa Cytosolic Phospholipase A2

1996 ◽  
Vol 271 (32) ◽  
pp. 19225-19231 ◽  
Author(s):  
Richard T. Pickard ◽  
X. Grace Chiou ◽  
Beth A. Strifler ◽  
Michael R. DeFelippis ◽  
Paul A. Hyslop ◽  
...  
Keyword(s):  
Phospholipase A2 ◽  
Cytosolic Phospholipase A2 ◽  
Catalytic Function ◽  
Cytosolic Phospholipase
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