The interconversion of isoforms of seminal ribonuclease: modelling key intermediates and trypsin effects 1 1Edited by J. Thorton

Cubellis, Maria Vittoria; D’Alessio, Giuseppe

doi:10.1006/jmbi.2000.3985

Cited by 2 publications

(2 citation statements)

References 26 publications

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“…2B). Previous biochemical investigations (30) and simulation studies (31) have shown that the first step in the MϭM i MXM interconversion is the unfolding and displacement of the Nterminal arm from each BS-RNase subunit. Thus, the block imposed by the disulfides on the displacement of the N-terminal arm would block the conversion of MϭM into MXM completely.…”

Section: Design Of a Stable Variant Of The Mϭm Isoform Of Bs-mentioning

confidence: 99%

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

Russo¹,

Antignani²,

Giancola³

et al. 2002

Journal of Biological Chemistry

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Seminal RNase (BS-RNase), a ribonuclease from bovine seminal vesicles, is a homodimeric enzyme with a strong cytotoxic activity selective for tumor cells. It displays the unusual structural feature of existing in solution as an equilibrium mixture of two quaternary isoforms. The major one is characterized by the swap between subunits of their N-terminal ends, whereas the minor isoform shows no swap. The tendency of the two isolated isoforms to interconvert into each other has so far made it difficult to attribute the functional properties of BS-RNase to either isoform. Herein, molecular modeling and site-directed mutagenesis were used to engineer the refolding pathway of BS-RNase and obtain a stable variant of its non-swapping isoform. The protein was engineered with two extra disulfide bridges linking the N-terminal helix of each subunit to the main body of the same subunit. Purified as an active enzyme, the BS-RNase variant was found to be very resistant to thermal denaturation. Its functional characterization revealed that the lack of swapping has a negative effect on the cytotoxic activity of BS-RNase.

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Section: Design Of a Stable Variant Of The Mϭm Isoform Of Bs-mentioning

confidence: 99%

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

Russo¹,

Antignani²,

Giancola³

et al. 2002

Journal of Biological Chemistry

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show abstract

“…These results, together with the kinetics of the tryptic action on both dimeric forms, led to the identification of a relevant interconversion intermediate [92] which would be the actual substrate for trypsin. This intermediate features an Nterminal arm that is dislocated from the rest of the protein [92,93]. A scheme of the postulated dimerization process of BS-RNase is shown in Fig.…”

Section: The Swapping Between Mxm and M=m Forms Proceeds Via An Intermentioning

confidence: 99%

The Structural Determinants that Lead to the Formation of Particular Oligomeric Structures in the Pancreatic-Type Ribonuclease Family

Benito¹,

Laurents²,

Ribó

et al. 2008

CPPS

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Pancreatic-type ribonucleases are a family of RNA degrading enzymes that share different degrees of sequence identity but a very similar 3D-structure. The prototype of this family is bovine pancreatic ribonuclease or ribonuclease A. This enzyme has been the object of landmark work on the folding, stability, protein chemistry, catalysis, enzyme-substrate interaction and molecular evolution. In the recent years, the interest in the study of pancreatic-type ribonucleases has increased due to the involvement of some members of this family in special biological functions. In addition, dimeric and also higher oligomeric structures can be attained by the members of this family. The oligomers described structurally to date are mainly formed by 3D-domain swapping, a process which consists of the exchange of identical domains (i.e. identical structural elements, usually the N- and C-termini) between the subunits and is considered to be a mechanism for amyloid-type aggregate formation. This review compares the dimeric and oligomeric structures of different members of the pancreatic-type ribonuclease family which are able to acquire these structures, namely, bovine seminal ribonuclease, ribonuclease A and its human counterpart, human pancreatic ribonuclease. A specific focus is placed on what is known about the structural determinants that lead to the acquisition of a particular oligomeric structure and on the proposed mechanism of 3D-swapping.

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The interconversion of isoforms of seminal ribonuclease: modelling key intermediates and trypsin effects 1 1Edited by J. Thorton

Cited by 2 publications

References 26 publications

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

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

The Structural Determinants that Lead to the Formation of Particular Oligomeric Structures in the Pancreatic-Type Ribonuclease Family

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