Unfolding pathway in red kidney bean acid phosphatase is dependent on ligand binding.


Abstract

Structural basis for ligand-induced protein stabilization was investigated in the case of an acid phosphatase (red kidney bean purple acid phosphatase (KBPAP)) from red kidney bean. Phosphate, a physiological ligand, increases the stability against solvent denaturation by 3.5 kcal/mol. Generality of phosphate stabilization was shown by similar effects with other KBPAP ligands viz. adenosine 5'-O-(thiotriphosphate), a nonhydrolyzable ligand, and arsenate, an inhibitor. The dissociation constant of phosphate obtained from denaturation curves matches with the dissociation constant estimated by conventional methods. The guanidinium chloride-mediated denaturation of KBPAP was monitored by several structural and functional parameters viz. activity, tryptophan fluorescence, 8-anilinonaphthalene 1-sulfonic acid binding, circular dichroism, and size exclusion chromatography, in the presence and absence of 10 mm phosphate. In the presence of phosphate, profiles of all the parameters shift to a higher guanidinium chloride concentration. Noncoincidence of these profiles in the absence of phosphate indicates multistate unfolding pathway for KBPAP; however, in the presence of phosphate, KBPAP unfolds with a single intermediate. Based on the crystal structure, we propose that the Arg258 may have an important role to play in stabilization mediated by phosphate. Study holds ProTherm entries: 5240 Extra Details:

Submission Details

ID: 8WNXWjTo

Submitter: Connie Wang

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

Version: 1

Publication Details
Cashikar AG;Rao NM,J. Biol. Chem. (1996) Unfolding pathway in red kidney bean acid phosphatase is dependent on ligand binding. PMID:8617740
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
6HWR 2019-04-03 1.95 Red kidney bean purple acid phosphatase in complex with adenosine divanadate
2QFP 2008-10-14 2.2 Crystal structure of red kidney bean purple acid phosphatase in complex with fluoride
4DHL 2012-12-12 2.3 Crystal structure of red kidney bean purple acid phosphatase in complex with Maybridge fragment MO07123
4DSY 2012-09-19 2.3 Crystal structure of red kidney bean purple acid phosphatase in complex with Maybridge fragment CC24201
6G46 2018-09-26 2.4 Red kidney bean purple acid phosphatase in complex with 2-(Naphthalen-1-yl)thiazole-4-carboxylic acid
2QFR 2008-10-14 2.4 Crystal structure of red kidney bean purple acid phosphatase with bound sulfate
1KBP 1996-10-14 2.65 KIDNEY BEAN PURPLE ACID PHOSPHATASE
4KBP 1996-12-07 2.7 KIDNEY BEAN PURPLE ACID PHOSPHATASE
4DT2 2012-09-19 2.7 Crystal structure of red kidney bean purple acid phosphatase in complex with Maybridge fragment CC27209
3KBP 1996-12-07 3.0 KIDNEY BEAN PURPLE ACID PHOSPHATASE

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Fe(3+)-Zn(2+) purple acid phosphatase P80366 PPAF_PHAVU