Thermodynamic stability of the two isoforms of bovine seminal ribonuclease.


Abstract

Bovine seminal ribonuclease (BS-RNase) is a dimeric protein with two identical subunits linked by two disulfide bridges, each subunit showing 80% of sequence identity with pancreatic RNase A. BS-RNase exists in two different quaternary conformations in solution: the MxM form, in which each subunit exchanges its alpha-helical N-terminal segment with its partner, and the M=M form with no exchange. By differential scanning microcalorimetry (DSC), the denaturation of the two dimeric forms of BS-RNase was found to be more complex than a simple two-state process. Monomeric derivatives of the dimeric protein follow instead a simple two-state mechanism, but are distinctly less stable than RNase A. The three-state N if I if D denaturation process of the two quaternary isoforms was interpreted by identifying in the dimers a central highly structured core, enclosing the covalently bonded subunit interface, which unfolds only after the periphery (mainly the N-terminal peptide) unfolds. Circular dichroism spectra of the two forms in the far-ultraviolet region show large differences between the secondary structure of the isoforms and that of the native BS-RNase mixture at equilibrium. This has been attributed to the presence in the equilibrium mixture of intermediate forms with displaced and disordered N-terminal alpha-helical segments. Study holds ProTherm entries: 8307, 8308 Extra Details: disulfide bridges; dimeric forms; secondary structure;,alpha-helical segments

Submission Details

ID: Kb2Wb8FM4

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:35 p.m.

Version: 1

Publication Details
Giancola C;Del Vecchio P;De Lorenzo C;Barone R;Piccoli R;D'Alessio G;Barone G,Biochemistry (2000) Thermodynamic stability of the two isoforms of bovine seminal ribonuclease. PMID:10891077
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
1QWQ 2003-09-16 Solution structure of the monomeric N67D mutant of Bovine Seminal Ribonuclease
2LFJ 2012-02-08 Solution structure of the monomeric derivative of BS-RNase
1N1X 2003-08-26 1.45 Crystal Structure Analysis of the monomeric [S-carboxyamidomethyl-Cys31, S-carboxyamidomethyl-Cys32] Bovine seminal ribonuclease
3DJQ 2009-06-30 1.53 Bovine Seminal Ribonuclease- Uridine 5' diphosphate complex
3DJP 2009-06-30 1.6 Bovine Seminal Ribonuclease- Uridine 3' phosphate complex
3DJO 2009-06-30 1.6 Bovine Seminal Ribonuclease uridine 2' phosphate complex
3DJV 2009-06-30 1.6 Bovine Seminal Ribonuclease- cytidine 3' phosphate complex
1N3Z 2003-08-26 1.65 Crystal structure of the [S-carboxyamidomethyl-Cys31, S-carboxyamidomethyl-Cys32] monomeric derivative of the bovine seminal ribonuclease in the liganded state
3DJX 2009-06-30 1.69 Bovine Seminal Ribonuclease- cytidine 5' phosphate complex
1BSR 1993-10-31 1.9 BOVINE SEMINAL RIBONUCLEASE STRUCTURE AT 1.9 ANGSTROMS RESOLUTION
11BG 1999-11-05 1.9 A POTENTIAL ALLOSTERIC SUBSITE GENERATED BY DOMAIN SWAPPING IN BOVINE SEMINAL RIBONUCLEASE
1TQ9 2004-09-14 2.0 Non-covalent swapped dimer of Bovine Seminal Ribonuclease in complex with 2'-DEOXYCYTIDINE-2'-DEOXYADENOSINE-3',5'-MONOPHOSPHATE
11BA 1999-03-26 2.06 BINDING OF A SUBSTRATE ANALOGUE TO A DOMAIN SWAPPING PROTEIN IN THE COMPLEX OF BOVINE SEMINAL RIBONUCLEASE WITH URIDYLYL-2',5'-ADENOSINE
1R5C 2004-04-13 2.1 X-ray structure of the complex of Bovine seminal ribonuclease swapping dimer with d(CpA)
1R3M 2003-11-18 2.2 Crystal structure of the dimeric unswapped form of bovine seminal ribonuclease
1Y94 2004-12-28 2.2 Crystal structure of the G16S/N17T/P19A/S20A/N67D Variant Of Bovine seminal Ribonuclease
1Y92 2004-12-28 2.2 Crystal structure of the P19A/N67D Variant Of Bovine seminal Ribonuclease
3BCM 2008-02-12 2.25 Crystal Structure of The Unswapped Form of P19A/L28Q/N67D BS-RNase
3BCO 2008-02-12 2.25 Crystal Structure of The Swapped FOrm of P19A/L28Q/N67D BS-RNase
4N4C 2013-11-06 2.48 Crystal structure of the C-terminal swapped dimer of a Bovine seminal ribonuclease mutant
1R5D 2004-04-13 2.5 X-ray structure of bovine seminal ribonuclease swapping dimer from a new crystal form
3BCP 2008-02-12 2.57 Crystal Structure of The Swapped non covalent form of P19A/L28Q/N67D BS-RNase

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Seminal ribonuclease P00669 RNS_BOVIN