Multiple-state equilibrium unfolding of guanidino kinases.


Abstract

The denaturant-induced equilibrium unfolding of octameric mitochondrial creatine kinase, dimeric cytosolic muscle-type creatine kinase, and monomeric arginine kinase was investigated. Stable unfolding intermediates for all three enzymes were manifested by a strongly biphasic red shift of intrinsic protein fluorescence upon increasing denaturant concentrations. In the intermediate state, all proteins were monomeric and enzymatically inactive, but still retained a globular shape. Native tertiary structure interactions were largely disrupted, while at least 50% of the secondary structures were conserved, as suggested by near- and far-UV circular dichroism, respectively. A significantly increased surface hydrophobicity of the intermediate conformation, compared to both the native and the fully unfolded states, was observed by the binding of the hydrophobic fluorescent dye ANS. The observed properties agree formally with the definition of the molten globule state, but can be alternatively explained by a sequential unfolding of individual domains, involving a transient exposure of domain interfaces. Very similar unfolding profiles for all three proteins suggest that the formation of stable unfolding intermediates is not a consequence of the specific oligomeric structures of the CKs but rather due to a common, probably two-domain architecture of the guanidino kinase protomers. Study holds ProTherm entries: 5287, 5288, 5289, 5290 Extra Details: additive : EDTA(0.1 mM),

Submission Details

ID: oVpwWLwJ4

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:29 p.m.

Version: 1

Publication Details
Gross M;Lustig A;Wallimann T;Furter R,Biochemistry (1995) Multiple-state equilibrium unfolding of guanidino kinases. PMID:7654688
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
4AM1 2012-07-11 1.25 Crystal structure of the marine crustacean decapod shrimp (Litopenaeus vannamei) arginine kinase in the absence of substrate or ligands.
4BG4 2013-09-04 1.6 Crystal structure of Litopenaeus vannamei arginine kinase in a ternary analog complex with arginine, ADP-Mg and NO3
1U6R 2005-08-02 1.65 Transition state analog complex of muscle creatine kinase (R134K) mutant
4Q2R 2014-09-03 1.65 Structural Proteomics From Crude Native Rod Outer Segments
4BHL 2013-09-04 1.9 Crystal structure of Litopenaeus vannamei arginine kinase in binary complex with arginine
3DRB 2009-03-17 2.0 Crystal structure of Human Brain-type Creatine Kinase
3B6R 2008-11-04 2.0 Crystal structure of Human Brain-type Creatine Kinase
3DRE 2009-03-17 2.2 Crystal structure of Human Brain-type Creatine Kinase
1G0W 2001-02-07 2.3 CRYSTAL STRUCTURE OF BOVINE RETINAL CREATINE KINASE
2CRK 1999-01-13 2.35 MUSCLE CREATINE KINASE
1QK1 2000-04-11 2.7 CRYSTAL STRUCTURE OF HUMAN UBIQUITOUS MITOCHONDRIAL CREATINE KINASE
5ZHQ 2019-03-20 3.0 Crystal structure of Scylla paramamosain arginine kinase
1I0E 2003-04-01 3.5 CRYSTAL STRUCTURE OF CREATINE KINASE FROM HUMAN MUSCLE

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
90.6 Creatine kinase M-type P00565 KCRM_CHICK
95.8 Creatine kinase M-type P06732 KCRM_HUMAN
96.1 Creatine kinase M-type P05123 KCRM_CANLF
96.6 Creatine kinase M-type Q9XSC6 KCRM_BOVIN
96.6 Creatine kinase M-type Q5XLD3 KCRM_PIG
97.1 Creatine kinase M-type P00564 KCRM_RAT
97.1 Creatine kinase M-type P07310 KCRM_MOUSE
99.2 Creatine kinase M-type P00563 KCRM_RABIT
91.3 Creatine kinase U-type, mitochondrial P70079 KCRU_CHICK
97.9 Creatine kinase U-type, mitochondrial Q29577 KCRU_PIG
96.3 Creatine kinase U-type, mitochondrial P25809 KCRU_RAT
98.4 Creatine kinase U-type, mitochondrial Q9TTK8 KCRU_BOVIN
97.6 Creatine kinase U-type, mitochondrial P30275 KCRU_MOUSE
100.0 Creatine kinase U-type, mitochondrial P12532 KCRU_HUMAN
95.3 Creatine kinase B-type P07335 KCRB_RAT
95.3 Creatine kinase B-type Q04447 KCRB_MOUSE
96.6 Creatine kinase B-type P12277 KCRB_HUMAN
96.9 Creatine kinase B-type Q29594 KCRB_PIG
96.9 Creatine kinase B-type Q5EA61 KCRB_BOVIN
97.4 Creatine kinase B-type P05124 KCRB_CANLF
100.0 Creatine kinase B-type P00567 KCRB_RABIT
91.2 Arginine kinase P51545 KARG_PENJP
91.0 Arginine kinase Q004B5 KARG0_PENVA
91.6 Arginine kinase C7E3T4 KARG_PENMO
91.3 Arginine kinase B0FRF9 KARG_PENVA
91.6 Arginine kinase B1PVZ9 KARG_METEN
92.7 Arginine kinase Q9U9J4 KARG_CARMA
92.7 Arginine kinase C9EIP1 KARG_SCYSE
93.0 Arginine kinase Q9GYX1 KARG_PACMR
93.0 Arginine kinase H6VGI3 KARG0_SCYPA
93.8 Arginine kinase Q9NH49 KARG_CALSI
93.8 Arginine kinase Q9NH48 KARG_ERISI
95.2 Arginine kinase H6VGI2 KARG_PROCL
100.0 Arginine kinase P14208 KARG_HOMGA