5HTD

Recombinant bovine beta-lactoglobulin variant L1A/I2S with endogenous ligand (sBlgB#1)

Summary for 5HTD

• Entry DOI: 10.2210/pdb5htd/pdb
• Descriptor: Beta-lactoglobulin, MYRISTIC ACID (3 entities in total)
• Functional Keywords: lipocalin, transport protein
• Biological source: Bos taurus (Bovine)
• Cellular location: Secreted: P02754
• Total number of polymer chains: 1
• Total formula weight: 18461.39

Authors

Loch, J.I.,Bonarek, P.,Tworzydlo, M.,Polit, A.,Hawro, B.,Lach, A.,Ludwin, E.,Lewinski, K. (deposition date: 2016-01-26, release date: 2016-07-20, Last modification date: 2024-01-10)

Primary citation

Loch, J.I.,Bonarek, P.,Tworzydo, M.,Polit, A.,Hawro, B.,Lach, A.,Ludwin, E.,Lewinski, K.
Engineered beta-Lactoglobulin Produced in E. coli: Purification, Biophysical and Structural Characterisation.
Mol Biotechnol., 58:605-618, 2016

PubMed Abstract: Functional recombinant bovine β-lactoglobulin has been produced by expression in E. coli using an engineered protein gene and purified to homogeneity by applying a new protocol. Mutations L1A/I2S introduced into the protein sequence greatly facilitate in vivo cleavage of the N-terminal methionine, allowing correctly folded and soluble protein suitable for biochemical, biophysical and structural studies to be obtained. The use of gel filtration on Sephadex G75 at the last purification step enables protein without endogenous ligand to be obtained. The physicochemical properties of recombinant β-lactoglobulin such as CD spectra, ligand binding (n, K a, ΔH, TΔS, ΔG), chemical and thermal stability (ΔG D, C mid) and crystal structure confirmed that the protein obtained is almost identical to the natural one. The substitutions of N-terminal residues did not influence the binding properties of the recombinant protein so that the lactoglobulin produced and purified according to our protocol is a good candidate for further engineering and potential use in pharmacology and medicine.

PubMed: 27380951
DOI: 10.1007/s12033-016-9960-z

Experimental method

X-RAY DIFFRACTION (2.5 Å)