Reversible unfolding of pyridoxal kinase.


Abstract

The unfolding of brain pyridoxal kinase by guanidinium HCl has been investigated at equilibrium. The overall process was reversible as judged from the complete recovery of catalytic activity after removal of guanidinium HCl. Unfolding of pyridoxal kinase was monitored by circular dichroism and fluorescence spectroscopy. The steepness of the spectroscopic changes between 0.2 and 1.5 M guanidinium HCl, and the lack of any discernible plateau suggests that unfolding of the monomer is a cooperative process. A compact intermediate on the unfolding pathway of pyridoxal kinase could not be detected by the method of denaturant gel filtration. The fluorescent analogs of the substrates ATP and pyridoxal were used to assess differences in stability among the domains of the protein. Based on fluorescence and steady emission anisotropy results, it is postulated that the nucleotide domain is more stable than the pyridoxal domain of the kinase. Study holds ProTherm entries: 5222, 5223 Extra Details: catalytic activity; cooperative process; compact intermediate;,unfolding pathway; nucleotide domain; pyridoxal domain

Submission Details

ID: iAqeYrv83

Submitter: Connie Wang

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

Version: 1

Publication Details
Pineda T;Churchich JE,J. Biol. Chem. (1993) Reversible unfolding of pyridoxal kinase. PMID:8376382
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
1LHP 2003-02-11 2.1 Crystal Structure of Pyridoxal Kinase from Sheep Brain
1LHR 2003-02-11 2.6 Crystal Structure of Pyridoxal Kinase complexed with ATP
1YGK 2005-07-05 2.6 Crystal Structure of Pyridoxal Kinase in Complex with Roscovitine and Derivatives
1YGJ 2005-07-05 2.7 Crystal Structure of Pyridoxal Kinase in Complex with Roscovitine and Derivatives
1RFU 2004-04-27 2.8 Crystal structure of pyridoxal kinase complexed with ADP and PLP
1RFT 2004-04-27 2.8 Crystal structure of pyridoxal kinase complexed with AMP-PCP and pyridoxamine
1RFV 2004-04-27 2.8 Crystal structure of pyridoxal kinase complexed with ADP
1YHJ 2005-07-05 2.8 Crystal Structure of Pyridoxal Kinase in Complex with Roscovitine and Derivatives

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
98.4 Pyridoxal kinase Q0II59 PDXK_BOVIN
100.0 Pyridoxal kinase P82197 PDXK_SHEEP