Calorimetric investigation of thermal stability and ligand-binding characteristics of disulfide-bond-cleaved ribonuclease T1.


Abstract

A combination of differential titration calorimetry and differential scanning calorimetry was used to study the effect of disulfide bond cleavage and reaction with iodoacetamide of ribonuclease T1 on both the binding of nucleotides and the thermal stability of the free enzyme species. Although guanosine monophosphates still bind to the active site of the modified protein the transition temperature of unfolding and the transition enthalpy decrease drastically indicating a relatively loose structure. The calorimetric data presented in this study suggest a cooperative linkage between the site of the disulfide bonds, the ligand-binding site, and the general thermodynamic stability of the enzyme. Study holds ProTherm entries: 7173, 7174, 7175, 7176, 7177 Extra Details: microcalorimetry; disulfide bonds; cooperativity;,ligand binding; ribonuclease T1

Submission Details

ID: zxdTZXsk

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:33 p.m.

Version: 1

Publication Details
Haun MF;Wirth M;R├╝terjans H,Eur. J. Biochem. (1995) Calorimetric investigation of thermal stability and ligand-binding characteristics of disulfide-bond-cleaved ribonuclease T1. PMID:7851431
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
3BU4 1998-09-14T00:00:00+0000 1.77 RIBONUCLEASE T1 COMPLEX WITH 2'GMP
5GSP 1997-12-09T00:00:00+0000 1.8 RIBONUCLEASE T1/3'-GMP, 9 WEEKS
4BIR 1998-01-13T00:00:00+0000 1.7 RIBONUCLEASE T1: FREE HIS92GLN MUTANT
1FYS 2000-10-03T00:00:00+0000 2.0 Ribonuclease T1 V16C mutant
1I2F 2001-02-09T00:00:00+0000 1.95 Ribonuclease T1 V16A mutant, form II
1I0V 2001-01-30T00:00:00+0000 1.23 Ribonuclease T1 in complex with 2'GMP (form I crystal)
1I0X 2001-01-30T00:00:00+0000 1.65 RIBONUCLEASE T1 IN COMPLEX WITH 2'GMP (FORM II CRYSTAL)
2GSP 1997-12-02T00:00:00+0000 1.8 RIBONUCLEASE T1/2',3'-CGPS AND 3'-GMP, 2 DAYS
1BIR 1996-01-04T00:00:00+0000 1.8 RIBONUCLEASE T1, PHE 100 TO ALA MUTANT COMPLEXED WITH 2' GMP
1I2E 2001-02-09T00:00:00+0000 1.8 Ribonuclease T1 V16A mutant, form I

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Guanyl-specific ribonuclease T1 P00651 RNT1_ASPOR