Cross‐strand disulphides in cell entry proteins: poised to act

Wouters, Merridee A.; Lau, Ken K.; Hogg, Philip J.

doi:10.1002/bies.10413

Cited by 63 publications

(101 citation statements)

References 51 publications

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“…To further characterize the rgp140 V3 intermonomer disulfide bond, we investigated its potential for PDI processing, and although we have not been able to assign an activity to the intersubunit disulfide bond, we have shown that it is potentially one of the disulfide bonds broken by PDI. This agrees with a hypothesis, based on the disulfide bonding pattern of gp120, that three disulfide bonds (spanning V1/V2, V3, and V4) are uncommon cross-strand disulfides (83). Such bonds are under stress and relatively unstable, making them prone to cleavage and release of their stored energy, which might drive conformational changes such as those involved in membrane fusion.…”

Section: Discussionsupporting

confidence: 91%

“…Such bonds are under stress and relatively unstable, making them prone to cleavage and release of their stored energy, which might drive conformational changes such as those involved in membrane fusion. With the V3 loop being involved in coreceptor binding, it was suggested that its disulfide bond would be the prime target for PDI processing (83).…”

Section: Discussionmentioning

confidence: 99%

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Stability of a Receptor-Binding Active Human Immunodeficiency Virus Type 1 Recombinant gp140 Trimer Conferred by Intermonomer Disulfide Bonding of the V3 Loop: Differential Effects of Protein Disulfide Isomerase on CD4 and Coreceptor Binding

Billington¹,

Hickling²,

Munro³

et al. 2007

J Virol

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Stable trimeric forms of human immunodeficiency virus recombinant gp140 (rgp140) are important templates for determining the structure of the glycoprotein to assist in our understanding of HIV infection and host immune response. Such information will aid the design of therapeutic drugs and vaccines. Here, we report the production of a highly stable and trimeric rgp140 derived from a HIV type 1 (HIV-1) subtype D isolate that may be suitable for structural studies. The rgp140 is functional in terms of binding to CD4 and three human monoclonal antibodies (17b, b12, and 2G12) that have broad neutralizing activities against a range of HIV-1 isolates from different subtypes. Treatment of rgp140 with protein disulfide isomerase (PDI) severely restricted 17b binding capabilities. The stable nature of the rgp140 was due to the lack of processing at the gp120/41 boundary and the presence of an intermonomer disulfide bond formed by the cysteines of the V3 loop. Further characterization showed the intermonomer disulfide bond to be a target for PDI processing. The relevance of these findings to the roles of the V3 domain and the timing of PDI action during the HIV infection process are discussed.

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Section: Discussionsupporting

confidence: 91%

Section: Discussionmentioning

confidence: 99%

Stability of a Receptor-Binding Active Human Immunodeficiency Virus Type 1 Recombinant gp140 Trimer Conferred by Intermonomer Disulfide Bonding of the V3 Loop: Differential Effects of Protein Disulfide Isomerase on CD4 and Coreceptor Binding

Billington¹,

Hickling²,

Munro³

et al. 2007

J Virol

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“…Although it is not yet known how often a single disulfide bond in vivo will be exposed to the force levels we explored in this study, it does seem likely that the sensitivity of any particular thiol͞disulfide exchange reaction to a pulling force will depend very specifically on the environment surrounding the bond as well as the type of chemical reaction involved. For example, ⌬x r is likely to depend on a number of factors that also affect the rate of the thiol͞disulfide exchange reaction, including the temperature (41), type of reducing agent (42), pH (42), electrostatics (43), the reaction mechanism (28), and the torsional strain present in the protein structure (21,44,45). Any combination of these effects that cause ⌬x r to be Ͼ1 Å would lead to a near 2-fold increase in reduction rate over just 20 pN of applied force, suggesting that force-catalyzed disulfide reduction may play an important role in vivo.…”

Section: Resultsmentioning

confidence: 99%

Force-dependent chemical kinetics of disulfide bond reduction observed with single-molecule techniques

Wiita

Ainavarapu²,

Huang³

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

328

372

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The mechanism by which mechanical force regulates the kinetics of a chemical reaction is unknown. Here, we use single-molecule force-clamp spectroscopy and protein engineering to study the effect of force on the kinetics of thiol͞disulfide exchange. Reduction of disulfide bonds through the thiol͞disulfide exchange chemical reaction is crucial in regulating protein function and is known to occur in mechanically stressed proteins. We apply a constant stretching force to single engineered disulfide bonds and measure their rate of reduction by DTT. Although the reduction rate is linearly dependent on the concentration of DTT, it is exponentially dependent on the applied force, increasing 10-fold over a 300-pN range. This result predicts that the disulfide bond lengthens by 0.34 Å at the transition state of the thiol͞disulfide exchange reaction. Our work at the single bond level directly demonstrates that thiol͞disulfide exchange in proteins is a force-dependent chemical reaction. Our findings suggest that mechanical force plays a role in disulfide reduction in vivo, a property that has never been explored by traditional biochemistry. Furthermore, our work also indicates that the kinetics of any chemical reaction that results in bond lengthening will be force-dependent.atomic force microscopy ͉ mechanochemistry T he intersection of force and chemistry has been studied for over a century, yet not much is known about this phenomenon compared with more common methods of chemical catalysis (1). There are a number of reasons for this discrepancy, but one of the most important factors remains that it is quite difficult to directly measure the effect of force on a bulk reaction. This difficulty arises because an applied force is not a scalar property of a system; it is associated with a vector. As a result, it is often not possible to directly probe the effect of force on a particular reaction because of heterogeneous application of force and a distribution of reaction orientations (1). To fully quantify the effect of an applied force on a chemical reaction, it is necessary to generate an experimental system where the reaction of interest is consistently oriented with respect to the applied force. Thus, recent advances in single-molecule techniques are particularly well suited to address this problem. The direct manipulation of single molecules allows for the application of force in a vector aligned with the reaction coordinate (2), avoiding the heterogeneity of bulk studies. Earlier works using singlemolecule techniques have described the rupture forces necessary to cleave single covalent bonds (reviewed in ref.

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“…A feature of ϪRHStaple bonds is the close proximity of the ␣-carbon atoms of the 2 cysteine residues that can impart strain on the bond. 27,30 The distance between the ␣-carbons of the Cys3-Cys5 bond is 4.01 Å (Table 2, average of the 2 structures), compared with a mean of 5.62 Å for all disulfides in a nonredundant set of X-ray structures. 31 The crossveinless 2 and procollagen 2A C domain structures were used to predict the structure of the VWF C2 domain using the I-TASSER protein structure prediction tool.…”

Section: Predicted Structure Of the C2 Domain Of Vwfmentioning

confidence: 99%

Lateral self-association of VWF involves the Cys2431-Cys2453 disulfide/dithiol in the C2 domain

Ganderton

Wong

Schroeder

et al. 2011

Blood

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VWF is a plasma protein that binds platelets to an injured vascular wall during thrombosis. When exposed to the shear forces found in flowing blood, VWF molecules undergo lateral self-association that results in a meshwork of VWF fibers. Fiber formation has been shown to involve thiol/disulfide exchange between VWF molecules. A C-terminal fragment of VWF was expressed in mammalian cells and examined for unpaired cysteine thiols using tandem mass spectrometry (MS). The VWF C2 domain Cys2431-Cys2453 disulfide bond was shown to be reduced in approximately 75% of the molecules. Fragments containing all 3 C domains or just the C2 domain formed monomers, dimers, and higher-order oligomers when expressed in mammalian cells. Mutagenesis studies showed that both the Cys2431-Cys2453 and nearby Cys2451-Cys2468 disulfide bonds were involved in oligomer formation. Our present findings imply that lateral VWF dimers form when a Cys2431 thiolate anion attacks the Cys2431 sulfur atom of the Cys2431-Cys2453 disulfide bond of another VWF molecule, whereas the Cys2451-Cys2468 disulfide/dithiol mediates formation of trimers and higher-order oligomers. These observations provide the basis for exploring defects in lateral VWF association in patients with unexplained hemorrhage or thrombosis. (Blood. 2011;118(19): 5312-5318) IntroductionVWF is a large, multimeric glycoprotein responsible for chaperoning the blood coagulation cofactor factor VIII and tethering platelets to the site of vascular endothelial injury. 1 It is synthesized by vascular endothelial cells and megakaryocytes and circulates as a series of multimers containing variable numbers of 500-kDa dimeric units. Circulating multimers range between 500 and 20 000 kDa in size, and although any size multimer is able to chaperone factor VIII, 2 it is the only the largest sizes that are able to effectively tether platelets. Each subunit contains binding sites for collagen and for platelet glycoproteins Ib and ␣IIb␤3. 1 Multimer size is regulated in the circulation. An excess of high-molecular-weight multimers can cause unwanted thrombosis and is associated with thrombotic thrombocytopenic purpura, 3 whereas a paucity of large multimers is associated with the bleeding disorder VWD. 1 Under normal conditions, the size of VWF multimers is controlled by ADAMTS13 (a disintegrin and metalloproteinase with thrombospondin type-1 motifs) proteolysis of the Tyr1605-Met1606 peptide bond in the A2 domain. 4 The tertiary and quaternary structure of VWF is influenced by the shear forces typically found in flowing blood. VWF undergoes a shear-dependent conformational change, 5 and individual molecules can self-associate under both static 6 and shear conditions. 7-9 VWF reversibly transitions from a loosely coiled ball to an elongated structure in response to shear. 5,10,11 Self-association of VWF has been reported in different systems. Soluble VWF perfused over a VWF-coated surface undergoes "homotypic" self-association that facilitates platelet adhesion. 9 In addition, VWF self-associates into ...

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Cross‐strand disulphides in cell entry proteins: poised to act

Cited by 63 publications

References 51 publications

Stability of a Receptor-Binding Active Human Immunodeficiency Virus Type 1 Recombinant gp140 Trimer Conferred by Intermonomer Disulfide Bonding of the V3 Loop: Differential Effects of Protein Disulfide Isomerase on CD4 and Coreceptor Binding

Stability of a Receptor-Binding Active Human Immunodeficiency Virus Type 1 Recombinant gp140 Trimer Conferred by Intermonomer Disulfide Bonding of the V3 Loop: Differential Effects of Protein Disulfide Isomerase on CD4 and Coreceptor Binding

Force-dependent chemical kinetics of disulfide bond reduction observed with single-molecule techniques

Lateral self-association of VWF involves the Cys2431-Cys2453 disulfide/dithiol in the C2 domain

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