Comparative analysis of refolding of chemically denatured beta-lactoglobulin types A and B using the dilution additive mode.


Abstract

The kinetic refolding of beta-lactoglobulin (BLG), types A and B, by beta-cyclodextrin, glucose and sorbitol has been investigated in aqueous solution using fluorescence, far UV-CD and UV-spectrophotometric techniques. A new Pd-complex has been used to denature the protein. CD and fluorescence studies indicated that when incubated with sugar, the denatured BLG is refolded into the native-like structure through the dilution additive mode resulting in a higher yield of active protein than without sugar. CD studies show that these sugars can induce a non-native alpha-helical structure in denatured BLG-A and -B, then aid in the refolding of the protein. Based on the present study, these sugars have a different effect on BLG-A than BLG-B because of their differences in protein thermal stability. BLG-A has a higher thermal stability than BLG-B due to differences in the amino acid sequences. Study holds ProTherm entries: 20574, 20575, 20576, 20577, 20578, 20579, 20580, 20581, 20582, 20583, 20584, 20585, 20586, 20587, 20588, 20589, 20590, 20591, 20592, 20593, 20594, 20595, 20596, 20597 Extra Details: beta-lactoglobulin; Sugar; Refolding; Thermal stability; alpha-helical structure

Submission Details

ID: JGjQZcMJ4

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:52 p.m.

Version: 1

Publication Details
Divsalar A;Saboury AA;Moosavi-Movahedi AA;Mansoori-Torshizi H,Int. J. Biol. Macromol. (2006) Comparative analysis of refolding of chemically denatured beta-lactoglobulin types A and B using the dilution additive mode. PMID:16417918
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
1CJ5 2000-04-26 BOVINE BETA-LACTOGLOBULIN A
1DV9 2000-02-09 STRUCTURAL CHANGES ACCOMPANYING PH-INDUCED DISSOCIATION OF THE B-LACTOGLOBULIN DIMER
4CK4 2014-10-08 1.12 Ovine beta-Lactoglobulin at Atomic Resolution
4TLJ 2014-06-18 1.17 Ultra-high resolution crystal structure of caprine Beta-lactoglobulin
4NLJ 2014-03-12 1.4 Crystal structure of sheep beta-lactoglobulin (space group P1)
4GNY 2013-02-13 1.64 Bovine beta-lactoglobulin complex with dodecyl sulfate
5NUK 2018-04-04 1.7 Engineered beta-lactoglobulin: variant I56F-L39A-M107F in complex with chlorpromazine (LG-FAF-CLP)
1BEB 1997-05-15 1.8 BOVINE BETA-LACTOGLOBULIN, LATTICE X
4KII 2013-09-04 1.85 Beta-lactoglobulin in complex with Cp*Rh(III)Cl N,N-di(pyridin-2-yl)dodecanamide
3UEV 2011-11-09 1.9 Bovine beta-lactoglobulin complex with myristic acid
4NLI 2014-03-12 1.9 Crystal structure of sheep beta-lactoglobulin (space group P3121)
3NQ3 2010-07-14 1.9 Bovine beta-lactoglobulin complex with capric acid
3NQ9 2010-07-14 1.9 Bovine beta-lactoglobulin complex with caprylic acid
4Y0S 2015-07-01 1.9 Goat beta-lactoglobulin complex with pramocaine (GLG-PRM)
6GHH 2018-06-20 1.9 Thermodynamic, Crystallographic and Computational Studies of Non Mammalian Fatty Acid Binding to Bovine b-Lactoglobulin
5NUN 2018-04-04 1.95 Engineered beta-lactoglobulin: variant F105L-L39A-M107F in complex with chlorpromazine (LG-LAF-CLP)
1B8E 2001-05-02 1.95 HIGH RESOLUTION CRYSTAL STRUCTURE OF THE BOVINE BETA-LACTOGLOBULIN (ISOFORMS A AND B) IN ORTHOROMBIC SPACE GROUP
5EEE 2016-11-09 1.95 Bovine lactoglobulin complex with decanol
1UZ2 2004-05-20 1.95 The Cys121Ser Mutant of Beta-Lactoglobulin
6QPD 2019-02-27 2.0 Engineered beta-lactoglobulin: variant I56F
5EED 2016-10-26 2.0 Ovine lactoglobulin complex with decanol
6GFS 2018-06-20 2.0 Thermodynamic, Crystallographic and Computational Studies of Non Mammalian Fatty Acid Binding to Bovine b-Lactoglobulin
6QI6 2019-01-30 2.0 Trigonal form of WT recombinant bovine beta-lactoglobulin
6FXB 2018-05-23 2.0 Bovine beta-lactoglobulin variant A at pH 4.0
1QG5 2001-04-21 2.0 HIGH RESOLUTION CRYSTAL STRUCTURE OF THE BOVINE BETA-LACTOGLOBULIN (ISOFORM A)
3UEW 2011-11-09 2.0 Bovine beta-lactoglobulin complex with palmitic acid
3KZA 2010-12-22 2.0 Crystal structure of Gyuba, a patched chimera of b-lactglobulin
4Y0Q 2015-07-01 2.0 Bovine beta-lactoglobulin complex with pramocaine crystallized from sodium citrate (BLG-PRM1)
6RWQ 2019-06-19 2.05 Engineered beta-lactoglobulin: variant F105L in complex with myristic acid
3UEU 2011-11-09 2.1 Bovine beta-lactoglobulin complex with lauric acid
4DQ4 2012-02-29 2.1 Bovine beta-lactoglobulin complex with linoleic acid
6GF9 2018-06-20 2.1 Thermodynamic, Crystallographic and Computational Studies of Non Mammalian Fatty Acid Binding to Bovine b-Lactoglobulin
6RWR 2019-06-19 2.1 Engineered beta-lactoglobulin: variant M107L in complex with myristic acid
2Q2M 2008-02-26 2.1 Beta-lactoglobulin (native)
6RWP 2019-06-19 2.1 Engineered beta-lactoglobulin: variant L58F in complex with myristic acid
4DQ3 2012-02-29 2.1 Bovine beta-lactoglobulin complex with oleic acid
6RYT 2019-06-26 2.1 Engineered beta-lactoglobulin: variant M107L
3UEX 2011-11-09 2.1 Bovine beta-lactoglobulin complex with stearic acid
6RYS 2019-06-26 2.15 Engineered beta-lactoglobulin: variant F105L
4IB7 2013-06-26 2.2 Bovine beta-lactoglobulin (isoform A) in complex with dodecyltrimethylammonium (DTAC)
4Y0P 2015-07-01 2.2 Bovine beta-lactoglobulin complex with tetracaine (BLG-TET)
6QPE 2019-02-27 2.2 Engineered beta-lactoglobulin: variant L58F
3NPO 2010-07-14 2.2 Bovine beta lactoglobulin unliganded form
4IB9 2013-06-26 2.2 Bovine beta-lactoglobulin (isoform B) in complex with dodecyltrimethylammonium (DTAC)
4IB6 2013-06-26 2.2 Bovine beta-lactoglobulin (isoform A) in complex with lauric acid (C12)
1BSQ 1998-09-02 2.22 STRUCTURAL AND FUNCTIONAL CONSEQUENCES OF POINT MUTATIONS OF VARIANTS A AND B OF BOVINE BETA-LACTOGLOBULIN
1BSO 1999-09-02 2.23 12-BROMODODECANOIC ACID BINDS INSIDE THE CALYX OF BOVINE BETA-LACTOGLOBULIN
1BSY 1999-01-27 2.24 STRUCTURAL BASIS OF THE TANFORD TRANSITION OF BOVINE BETA-LACTOGLOBULIN FROM CRYSTAL STRUCTURES AT THREE PH VALUES; PH 7.1
4Y0R 2015-07-01 2.3 Bovine beta-lactoglobulin complex with pramocaine crystallized from ammonium sulphate (BLG-PRM2)
6GE7 2018-06-20 2.3 Thermodynamic, Crystallographic and Computational Studies of Non Mammalian Fatty Acid Binding to Bovine b-Lactoglobulin
6NKQ 2019-01-23 2.3 The structure of bovine beta-lactoglobulin in novel crystals grown at pH 3.8
4IBA 2013-06-26 2.3 Bovine beta-lactoglobulin (isoform B) in complex with dodecyl sulphate (SDS)
4OMX 2014-11-19 2.3 Crystal structure of goat beta-lactoglobulin (trigonal form)
4IB8 2013-06-26 2.3 Bovine beta-lactoglobulin (isoform A) in complex with dodecyl sulphate (SDS)
5NUM 2018-04-04 2.3 Engineered beta-lactoglobulin: variant F105L-L39A in complex with chlorpromazine (LG-LA-CLP)
4OMW 2014-11-19 2.3 Crystal structure of goat beta-lactoglobulin (orthorhombic form)
1GX9 2002-06-13 2.34 BOVINE BETA-LACTOGLOBULIN COMPLEXED WITH RETINOIC ACID, TRIGONAL LATTICE Z
1GXA 2002-06-13 2.35 BOVINE BETA-LACTOGLOBULIN COMPLEXED WITH RETINOL AND PALMITIC ACID, TRIGONAL LATTICE Z
3PH5 2011-09-21 2.4 Bovine beta lactoglobulin crystallized through ligandation of yttrium cations
5HTE 2016-07-20 2.4 Recombinant bovine beta-lactoglobulin variant L1A/I2S (sBlgB#2)
4LZU 2014-08-06 2.4 Bovine beta-lactoglobulin crystallized in the presence of 2 mM zinc chloride
2GJ5 2007-10-02 2.4 Crystal structure of a secondary vitamin D3 binding site of milk beta-lactoglobulin
1GX8 2002-06-13 2.4 BOVINE BETA-LACTOGLOBULIN COMPLEXED WITH RETINOL, TRIGONAL LATTICE Z
4LZV 2014-08-06 2.44 Bovine beta-lactoglobulin crystallized in the presence of 20 mM zinc chloride
2BLG 1999-01-27 2.46 STRUCTURAL BASIS OF THE TANFORD TRANSITION OF BOVINE BETA-LACTOGLOBULIN FROM CRYSTAL STRUCTURES AT THREE PH VALUES; PH 8.2
5LKF 2016-08-17 2.5 Bovine beta-lactoglobulin complex with myristic acid at high pressure (0.55 GPa)
5HTD 2016-07-20 2.5 Recombinant bovine beta-lactoglobulin variant L1A/I2S with endogenous ligand (sBlgB#1)
2Q39 2008-02-26 2.5 Beta-lactoglobulin (low humidity)
5K06 2016-07-20 2.5 Recombinant bovine beta-lactoglobulin with uncleaved N-terminal methionine (rBlgB)
1B0O 1999-02-02 2.5 BOVINE BETA-LACTOGLOBULIN COMPLEXED WITH PALMITATE, LATTICE Z
6QI7 2019-01-30 2.5 Engineered beta-lactoglobulin: variant L39Y in complex with endogenous ligand
3PH6 2011-09-21 2.53 Bovine beta lactoglobulin crytsallized through ligandation of yttrium
3BLG 1999-01-27 2.56 STRUCTURAL BASIS OF THE TANFORD TRANSITION OF BOVINE BETA-LACTOGLOBULIN FROM CRYSTAL STRUCTURES AT THREE PH VALUES; PH 6.2
5NUJ 2018-04-04 2.6 Engineered beta-lactoglobulin: variant I56F-L39A in complex with chlorpromazine (LG-FA-CLP)
5Y5C 2018-08-08 2.61 Crystal structure of dha bind to bovine beta-lactoglobulin
5LKE 2016-08-10 2.8 Bovine beta-lactoglobulin complex with myristic acid, ambient pressure
2R56 2007-11-13 2.8 Crystal Structure of a Recombinant IgE Fab Fragment in Complex with Bovine Beta-Lactoglobulin Allergen
5IO5 2016-03-16 2.85 Unliganded form of bovine beta-lactoglobulin, ambient pressure
5IO7 2016-03-16 2.85 Bovine beta-lactoglobulin complex with dodecane at high pressure (0.43 GPa)
2Q2P 2008-02-26 2.96 Beta-lactoglobulin (reverse native)
2AKQ 2005-08-16 3.0 The structure of bovine B-lactoglobulin A in crystals grown at very low ionic strength

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
95.7 Beta-lactoglobulin P67975 LACB_OVIMU
95.7 Beta-lactoglobulin P67976 LACB_SHEEP
96.3 Beta-lactoglobulin P02756 LACB_CAPHI
98.8 Beta-lactoglobulin P02755 LACB_BUBBU
100.0 Beta-lactoglobulin P02754 LACB_BOVIN