7ZLF

Mutant L39Y-L58F of recombinant bovine beta-lactoglobulin in complex with endogenous fatty acid

This is a non-PDB format compatible entry.

Summary for 7ZLF

• Entry DOI: 10.2210/pdb7zlf/pdb
• Related: 7Z9Z 7ZA0 7ZDC
• Descriptor: Beta-lactoglobulin, LAURIC ACID, 1,2-ETHANEDIOL, ... (6 entities in total)
• Functional Keywords: lactoglobulin, mutation, ligand, transport protein
• Biological source: Bos taurus (cattle)
• Total number of polymer chains: 1
• Total formula weight: 18773.02

Authors

Loch, J.I.,Kurpiewska, K.,Bonarek, P. (deposition date: 2022-04-14, release date: 2023-04-26, Last modification date: 2024-11-13)

Primary citation

Bonarek, P.,Mularczyk, D.,Loch, J.I.,Kurpiewska, K.,Dziedzicka-Wasylewska, M.
beta-Lactoglobulin variants as potential carriers of pramoxine: Comprehensive structural and biophysical studies.
J.Mol.Recognit., 36:e3052-e3052, 2023

PubMed Abstract: β-Lactoglobulin (BLG) is a member of the lipocalin family. As other proteins from this group, BLG can be modified to bind specifically compounds of medical interests. The aim of this study was to evaluate the role of two mutations, L39Y and L58F, in the binding of topical anesthetic pramoxine (PRM) to β-lactoglobulin. Circular dichroism spectroscopy, isothermal titration calorimetry (ITC), and X-ray crystallography were used to understand the mechanisms of BLG-PRM interactions. Studies were performed for three new BLG mutants: L39Y, L58F, and L39Y/L58F. ITC measurements indicated a significant increase in the affinity to the PRM of variants L58F and L39Y. Measurements taken for the double mutant L39Y/L58F showed the additivity of two mutations leading to about 80-fold increase in the affinity to PRM in comparison to natural protein BLG from bovine milk. The determined crystal structures revealed that pramoxine is accommodated in the β-barrel interior of BLG mutants and stabilized by hydrophobic interactions. The observed additive effect of two mutations on drug binding opens the possibility for further designing of new BLG variants with high affinity to selected drugs.

PubMed: 37610054
DOI: 10.1002/jmr.3052

Experimental method

X-RAY DIFFRACTION (2.1 Å)