Engineering the Refolding Pathway and the Quaternary Structure of Seminal Ribonuclease by Newly Introduced Disulfide Bridges

Russo, Aniello; Antignani, Antonella; Giancola, Concetta; D’Alessio, Giuseppe

doi:10.1074/jbc.m207141200

Cited by 4 publications

(7 citation statements)

References 47 publications

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“…On the basis of H R -H R (i,i+1) and HN-H R (i,i+1) NOEs, we assigned cis isomerism to proline residues 93 and 114, whereas trans isomerism was observed for Pro 117 and Pro 42 , on the basis of H R -H δ (i,i+1) and HN-H δ (i,i+1) NOEs, consistent with the findings for RNase A (38,39) and BS-RNase (7,40). Pro 19 , which is not present in RNase A, was also assigned a trans configuration on the basis of a NOE between Ser 18 H R and Pro 19 (30), transferred to the program package DYANA 1.5 (41), and converted into upper distance limits by using CALIBA (42). Spin coupling values were input into HABAS (43) to obtain the possible ranges of torsion angles with simultaneous consideration of the NOE-derived distance restraints.…”

supporting

confidence: 86%

“…Domain swapping in BS-RNase was reported to be determinant for some special, noncatalytic, biological actions, such as allostery (17), cytotoxicity toward malignant cells (18,19), and immunosuppression and antispermatogenesis (20), which may support a physiological role. The active sites of the MxM form are located at the two interfaces between the swapping tails and the main protein bodies and are formed by residues belonging to different chains; in turn, each chain is involved in both active sites.…”

mentioning

confidence: 99%

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The Swapping of Terminal Arms in Ribonucleases: Comparison of the Solution Structure of Monomeric Bovine Seminal and Pancreatic Ribonucleases

et al. 2003

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Bovine seminal ribonuclease (BS-RNase), the only dimeric protein among the pancreatic-like ribonucleases, is endowed with special structural features and with biological functions beyond enzymatic activity. In solution, the protein exists as an equilibrium mixture of two forms, with or without exchange (or swapping) of the N-terminal arms. After selective reduction and alkylation of the two intrachain disulfide bridges, the dimeric protein can be transformed into a monomeric derivative that has a ribonuclease activity higher than that of the parent dimeric protein but is devoid of the special biological functions. A detailed investigation of the structural features of this protein in solution, in comparison with those of other monomeric ribonucleases, may help unveil the structural details which induce swapping of the N-terminal arms of BS-RNase. The solution structure of the recombinant monomeric form of BS-RNase, as determined by 3D heteronuclear NMR, shows close similarity with that of bovine pancreatic ribonuclease (RNase A) in all regions characterized by regular elements of secondary structure. However, significant differences are present in the flexible regions, which could account for the different behavior of the two proteins. To characterize in detail these regions, we have measured H/D exchange rate constants, temperature coefficients and heteronuclear NOEs of backbone amides for both RNase A and monomeric BS-RNase. The results indicate a large difference in the backbone flexibility of the hinge peptide segment 16-22 of the two proteins, which could provide the molecular basis to explain the ability of BS-RNase subunits to swap their N-terminal arms.

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confidence: 86%

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confidence: 99%

The Swapping of Terminal Arms in Ribonucleases: Comparison of the Solution Structure of Monomeric Bovine Seminal and Pancreatic Ribonucleases

et al. 2003

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“…Within the MXM form, an exchange of N -terminal -helices between subunits occurs whereas in the M=M form this exchange is not observed [92,95,96]. The absence of Nterminal swapping in the M=M enzyme may account for the differences in conformational stability, enzymatic and biological action of these two isoforms [6,75,93,95,98]. While both isoforms and monomeric BS-RNase exhibit enzymatic activity only MXM-BS-RNase exhibits significant cytotoxic and antitumor effects in cultured malignant cells and solid tumors [91,98].…”

Section: Bovine Seminal-rnase (Bs-rnase)mentioning

confidence: 98%

“…The absence of Nterminal swapping in the M=M enzyme may account for the differences in conformational stability, enzymatic and biological action of these two isoforms [6,75,93,95,98]. While both isoforms and monomeric BS-RNase exhibit enzymatic activity only MXM-BS-RNase exhibits significant cytotoxic and antitumor effects in cultured malignant cells and solid tumors [91,98]. The cytotoxicity of MXM-BS-RNase is believed to be the result of its inherent RNase activity, its ability to evade inhibition by cellular RNase inhibitors and its structural stability within the reducing environment of the cytosol [93].…”

Section: Bovine Seminal-rnase (Bs-rnase)mentioning

confidence: 99%

Destroying RNA as a Therapeutic Approach

Tafech¹,

Bassett²,

Sparanese³

et al. 2006

CMC

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The ability to target RNA, mRNA and viral RNA in particular, for degradation is a powerful approach in molecular biology and pharmacology. Such approaches can be used in the study of gene function as in functional genomics, in the identification of disease-associated genes, and for the treatment of human diseases. This review provides a comprehensive up-to-date look at all the current available technologies used for the destruction of RNA, with a focus on their therapeutic potential. This includes approaches that utilize the activity of protein ribonucleases such as antisense oligonucleotide, small interfering RNA, RNase P-associated external guide sequence, onconase and bovine seminal RNase. Sequence-specific approaches that do not utilize activity of protein ribonucleases, such as ribozyme and DNazyme, are also reviewed and discussed. This review should provide a useful starting framework for researchers interested in using the RNA-destruction methodologies on the bench and in the clinic, and serves as a stimulus for further development of novel and more potent RNA degradation technologies. This is particularly critical, given the anticipation of discoveries of new cellular RNA degradation machineries and human diseases that are associated with dysfunctional RNA molecules.

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“…This M×M/M = M equilibrium has been deeply investigated (282) and partially affects the catalytic activity of the enzyme (278). The role of the inter-subunit disulfides has also been analyzed (283, 284), as well as the one of the hinge loop connecting the N-terminus with the protein core. In addition, P19 and L28, as well as R80 residues, have been shown to influence the M = M vs. M×M equilibrium (285–287).…”

Section: Rnase Oligomerization: a Strategy To Obtain Stable And More mentioning

confidence: 99%

Biological Activities of Secretory RNases: Focus on Their Oligomerization to Design Antitumor Drugs

Gotte

Menegazzi

2019

Front. Immunol.

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Ribonucleases (RNases) are a large number of enzymes gathered into different bacterial or eukaryotic superfamilies. Bovine pancreatic RNase A, bovine seminal BS-RNase, human pancreatic RNase 1, angiogenin (RNase 5), and amphibian onconase belong to the pancreatic type superfamily, while binase and barnase are in the bacterial RNase N1/T1 family. In physiological conditions, most RNases secreted in the extracellular space counteract the undesired effects of extracellular RNAs and become protective against infections. Instead, if they enter the cell, RNases can digest intracellular RNAs, becoming cytotoxic and having advantageous effects against malignant cells. Their biological activities have been investigated either in vitro, toward a number of different cancer cell lines, or in some cases in vivo to test their potential therapeutic use. However, immunogenicity or other undesired effects have sometimes been associated with their action. Nevertheless, the use of RNases in therapy remains an appealing strategy against some still incurable tumors, such as mesothelioma, melanoma, or pancreatic cancer. The RNase inhibitor (RI) present inside almost all cells is the most efficacious sentry to counteract the ribonucleolytic action against intracellular RNAs because it forms a tight, irreversible and enzymatically inactive complex with many monomeric RNases. Therefore, dimerization or multimerization could represent a useful strategy for RNases to exert a remarkable cytotoxic activity by evading the interaction with RI by steric hindrance. Indeed, the majority of the mentioned RNases can hetero-dimerize with antibody derivatives, or even homo-dimerize or multimerize, spontaneously or artificially. This can occur through weak interactions or upon introducing covalent bonds. Immuno-RNases, in particular, are fusion proteins representing promising drugs by combining high target specificity with easy delivery in tumors. The results concerning the biological features of many RNases reported in the literature are described and discussed in this review. Furthermore, the activities displayed by some RNases forming oligomeric complexes, the mechanisms driving toward these supramolecular structures, and the biological rebounds connected are analyzed. These aspects are offered with the perspective to suggest possible efficacious therapeutic applications for RNases oligomeric derivatives that could contemporarily lack, or strongly reduce, immunogenicity and other undesired side-effects.

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Engineering the Refolding Pathway and the Quaternary Structure of Seminal Ribonuclease by Newly Introduced Disulfide Bridges

Cited by 4 publications

References 47 publications

The Swapping of Terminal Arms in Ribonucleases: Comparison of the Solution Structure of Monomeric Bovine Seminal and Pancreatic Ribonucleases

The Swapping of Terminal Arms in Ribonucleases: Comparison of the Solution Structure of Monomeric Bovine Seminal and Pancreatic Ribonucleases

Destroying RNA as a Therapeutic Approach

Biological Activities of Secretory RNases: Focus on Their Oligomerization to Design Antitumor Drugs

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