Anion-induced stabilization of human serum albumin prevents the formation of intermediate during urea denaturation.


Abstract

The unfolding of human serum albumin (HSA), a multidomain protein, by urea was followed by far-UV circular dichroism (CD), intrinsic fluorescence, and ANS fluorescence measurements. The urea-induced transition, which otherwise was a two-step process with a stable intermediate at around 4.8 M urea concentration as monitored by far-UV CD and intrinsic fluorescence, underwent a single-step cooperative transition in the presence of 1.0 M KCl. The free energy of stabilization (DeltaDelta G(H2O)D) in the presence of 1 M KCl was found to be 1,090 and 1,200 cal/mol as determined by CD and fluorescence, respectively. The salt stabilization occurred in the first transition (0-5.0 M urea), which corresponded to the formation of intermediate (I) state from the native (N) state, whereas the second transition, corresponding to the unfolding of I state to denatured (D) state, remained unaffected. Urea denaturation of HSA as monitored by tryptophan fluorescence of the lone tryptophan residue (Trp(214)) residing in domain II of the protein, followed a single-step transition suggesting that domain(s) I and/or III is (are) involved in the intermediate formation. This was also confirmed by the acrylamide quenching of tryptophan fluorescence at 5 M urea, which exhibited little change in the value of Stern-Volmer constant. ANS fluorescence data also showed single-step transition reflecting the absence of accumulation of hydrophobic patches. The stabilizing potential of various salts studied by far-UV CD and intrinsic fluorescence was found to follow the order: NaClO(4) > NaSCN >Na(2)SO(4) >KBr >KCl >KF. A comparison of the effects of various potassium salts revealed that anions were chiefly responsible in stabilizing HSA. The above series was found similar to the electroselectivity series of anions towards the anion-exchange resins and reverse of the Hofmeister series, suggesting that preferential binding of anions to HSA rather than hydration, was primarily responsible for stabilization. Further, single-step transition observed with GdnHCl can be ascribed to its ionic character as the free energy change associated with urea denaturation in the presence of 1.0 M KCl (5,980 cal/mol) was similar to that obtained with GdnHCl (5,870 cal/mol). Study holds ProTherm entries: 8043, 8044, 8045, 8046 Extra Details: the transition is from native to intermediate N->I

Submission Details

ID: rGJS6RsX4

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:35 p.m.

Version: 1

Publication Details
Muzammil S;Kumar Y;Tayyab S,Proteins (2000) Anion-induced stabilization of human serum albumin prevents the formation of intermediate during urea denaturation. PMID:10813828
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
1YSX 2005-06-07 Solution structure of domain 3 from human serum albumin complexed to an anti-apoptotic ligand directed against Bcl-xL and Bcl-2
2N0X 2015-04-29 Three dimensional structure of EPI-X4, a human albumin-derived peptide that regulates innate immunity through the CXCR4/CXCL12 chemotactic axis and antagonizes HIV-1 entry
2ESG 2006-01-31 Solution structure of the complex between immunoglobulin IgA1 and human serum albumin
6HSC 2019-10-23 1.9 Structure of Human Serum Albumin in complex with Aristolochic Acid at 1.9 A resolution
1N5U 2003-06-24 1.9 X-RAY STUDY OF HUMAN SERUM ALBUMIN COMPLEXED WITH HEME
6QIO 2020-02-05 1.95 Ternary complex of FcRn ectodomain, FcRn binding optimised human serum albumin and the human growth hormone derivative somapacitan
4L8U 2013-07-24 2.01 X-ray study of human serum albumin complexed with 9 amino camptothecin
3SQJ 2011-11-02 2.05 Recombinant human serum albumin from transgenic plant
5Z0B 2018-12-19 2.17 Crystal structure of plasma-derived human serum albumin
4Z69 2016-01-27 2.19 Human serum albumin complexed with palmitic acid and diclofenac
3A73 2010-01-19 2.19 Crystal Structure Analysis of Human serum albumin complexed with delta 12-prostaglandin J2
1E7A 2001-01-17 2.2 Crystal structure of human serum albumin complexed with the general anesthetic propofol
3UIV 2012-11-07 2.2 Human serum albumin-myristate-amantadine hydrochloride complex
4G03 2013-07-10 2.22 High-resolution Crystal Structural Variance Analysis between Recombinant and Wild-type Human Serum Albumin
2BXH 2005-09-22 2.25 Human serum albumin complexed with indoxyl sulfate
5ID7 2016-05-11 2.26 Crystal structure of human serum albumin in complex with phosphorodithioate derivative of myristoyl cyclic phosphatidic acid (cPA)
6A7P 2018-10-24 2.28 Human serum albumin complexed with aripiprazole
4G04 2013-07-10 2.3 High-resolution Crystal Structural Variance Analysis between Recombinant and Wild-type Human Serum Albumin
2BXP 2005-09-22 2.3 Human serum albumin complexed with myristate and phenylbutazone
4EMX 2012-04-25 2.3 Crystal structure analysis of Human Serum Albumin in complex with chloride anions at cryogenic temperature
3JRY 2010-04-21 2.3 Human Serum albumin with bound Sulfate
4E99 2012-06-06 2.3 Human Serum Albumin Complex with Perfluorooctane Sulfonate Potassium
4HGM 2013-05-08 2.34 Shark IgNAR Variable Domain
4BKE 2014-02-05 2.35 Recombinant human serum albumin with palmitic acid. Synthetic cationic antimicrobial peptides bind with their hydrophobic parts to drug site II of human serum albumin
2BXA 2005-09-22 2.35 Human serum albumin complexed with 3-carboxy-4-methyl-5-propyl-2- furanpropanoic acid (CMPF)
1E7B 2001-01-17 2.38 Crystal structure of human serum albumin complexed with the general anesthetic halothane
6EZQ 2017-12-13 2.39 human Serum Albumin complexed with NBD-C12 fatty acid
4K71 2013-10-23 2.4 Crystal structure of a high affinity Human Serum Albumin variant bound to the Neonatal Fc Receptor
2BXK 2005-09-22 2.4 Human serum albumin complexed with myristate, azapropazone and indomethacin
1E7C 2001-01-17 2.4 HUMAN SERUM ALBUMIN COMPLEXED WITH MYRISTIC ACID and the general anesthetic halothane
1GNI 2002-01-01 2.4 HUMAN SERUM ALBUMIN COMPLEXED WITH cis-9-OCTADECENOIC ACID (OLEIC ACID)
1HK4 2003-05-16 2.4 HUMAN SERUM ALBUMIN COMPLEXED WITH THYROXINE (3,3',5,5'-TETRAIODO-L-THYRONINE) and myristic acid (tetradecanoic acid)
2XW0 2010-11-10 2.4 Human serum albumin complexed with dansyl-L-phenylalanine
4LA0 2013-07-24 2.4 X-ray study of human serum albumin complexed with bicalutamide
4L9K 2013-07-24 2.4 X-ray study of human serum albumin complexed with camptothecin
2XVV 2010-11-10 2.4 Human serum albumin complexed with dansyl-L-asparagine and myristic acid
4IW2 2013-04-24 2.41 HSA-glucose complex
2VUE 2008-06-17 2.42 Human serum albumin complexed with 4Z,15E-bilirubin-IX-alpha
1E7H 2000-12-08 2.43 HUMAN SERUM ALBUMIN COMPLEXED WITH HEXADECANOIC ACID (PALMITIC ACID)
1E7F 2000-11-06 2.43 HUMAN SERUM ALBUMIN COMPLEXED WITH DODECANOIC ACID (LAURIC ACID)
6QIP 2020-02-05 2.45 Ternary complex of FcRn ectodomain, FcRn binding optimised human serum albumin and the albumin-biniding side chain of the human growth hormone derivative somapacitan
2BXI 2005-09-22 2.5 Human serum albumin complexed with myristate and azapropazone
1BM0 1999-07-28 2.5 CRYSTAL STRUCTURE OF HUMAN SERUM ALBUMIN
1BJ5 1998-11-04 2.5 HUMAN SERUM ALBUMIN COMPLEXED WITH MYRISTIC ACID
1H9Z 2001-06-20 2.5 Human Serum Albumin Complexed With Myristic Acid and the R-(+) enantiomer of warfarin
2BXM 2005-09-22 2.5 Human serum albumin complexed with myristate and indomethacin
1E7E 2000-11-06 2.5 HUMAN SERUM ALBUMIN COMPLEXED WITH DECANOIC ACID (CAPRIC ACID)
1E7G 2000-11-06 2.5 Human serum albumin complexed with tetradecanoic acid (myristic acid)
1HA2 2001-06-20 2.5 Human Serum Albumin Complexed With Myristic Acid and the S-(-) enantiomer of warfarin
1AO6 1998-05-27 2.5 CRYSTAL STRUCTURE OF HUMAN SERUM ALBUMIN
2XW1 2010-11-10 2.5 Human serum albumin complexed with dansyl-L-norvaline
2VDB 2008-02-26 2.52 Structure of human serum albumin with S-naproxen and the GA module
5GIY 2017-07-19 2.54 HSA-Palmitic acid-[RuCl5(ind)]2-
4IW1 2013-04-24 2.56 HSA-fructose complex
2XVU 2010-11-10 2.6 Human serum albumin complexed with dansyl-L-asparagine
3B9L 2008-05-27 2.6 Human serum albumin complexed with myristate and AZT
1E78 2001-03-23 2.6 Crystal structure of human serum albumin
2BXO 2005-09-22 2.6 Human serum albumin complexed with myristate and oxyphenbutazone
2BXQ 2005-09-22 2.6 Human serum albumin complexed with myristate, phenylbutazone and indomethacin
3LU8 2010-10-27 2.6 Human serum albumin in complex with compound 3
5VNW 2018-02-21 2.6 Crystal structure of Nb.b201 bound to human serum albumin
1GNJ 2002-01-01 2.6 HUMAN SERUM ALBUMIN COMPLEXED WITH cis-5,8,11,14-EICOSATETRAENOIC ACID (ARACHIDONIC ACID)
2BXL 2005-09-22 2.6 Human serum albumin complexed with myristate and 3,5-diiodosalicylic acid
3TDL 2012-06-27 2.6 Structure of human serum albumin in complex with DAUDA
5YB1 2018-09-05 2.62 Structure and function of human serum albumin-metal agent complex
5YOQ 2018-02-07 2.65 Human serum albumin complexed with sodium 4-phenylbutyrate
1HK1 2003-05-16 2.65 HUMAN SERUM ALBUMIN COMPLEXED WITH THYROXINE (3,3',5,5'-TETRAIODO-L-THYRONINE)
2XVW 2010-11-10 2.65 Human serum albumin complexed with dansyl-L-arginine and myristic acid
5IJF 2016-03-16 2.65 Crystal structure of Human Serum Albumin in the presence of 0.5 mM zinc at pH 9.0
2BXN 2005-09-22 2.65 Human serum albumin complexed with myristate and iodipamide
3LU6 2010-10-27 2.7 Human serum albumin in complex with compound 1
5UJB 2017-05-03 2.7 Structure of a Mcl-1 Inhibitor Binding to Site 3 of Human Serum Albumin
4L9Q 2013-07-24 2.7 X-ray study of human serum albumin complexed with teniposide
4LB9 2013-07-24 2.7 X-ray study of human serum albumin complexed with etoposide
1HK5 2003-05-16 2.7 HUMAN SERUM ALBUMIN MUTANT R218H COMPLEXED WITH THYROXINE (3,3',5,5'-TETRAIODO-L-THYRONINE) and myristic acid (tetradecanoic acid)
2BX8 2005-09-22 2.7 Human serum albumin complexed with azapropazone
2I2Z 2006-12-12 2.7 Human serum albumin complexed with myristate and aspirin
2BXG 2005-09-22 2.7 Human serum albumin complexed with ibuprofen
1E7I 2000-11-06 2.7 HUMAN SERUM ALBUMIN COMPLEXED WITH OCTADECANOIC ACID (STEARIC ACID)
3B9M 2008-05-27 2.7 Human serum albumin complexed with myristate, 3'-azido-3'-deoxythymidine (AZT) and salicylic acid
1TF0 2004-09-07 2.7 Crystal structure of the GA module complexed with human serum albumin
2XSI 2010-11-10 2.7 Human serum albumin complexed with dansyl-L-glutamate and myristic acid
2YDF 2011-04-27 2.75 HUMAN SERUM ALBUMIN COMPLEXED WITH IOPHENOXIC ACID
1HK3 2003-05-16 2.8 Human serum albumin mutant r218p complexed with thyroxine (3,3',5,5'-tetraiodo-l-thyronine)
1UOR 1998-05-27 2.8 X-RAY STUDY OF RECOMBINANT HUMAN SERUM ALBUMIN. PHASES DETERMINED BY MOLECULAR REPLACEMENT METHOD, USING LOW RESOLUTION STRUCTURE MODEL OF TETRAGONAL FORM OF HUMAN SERUM ALBUMIN
3CX9 2009-04-28 2.8 Crystal Structure of Human serum albumin complexed with Myristic acid and lysophosphatidylethanolamine
3LU7 2010-10-27 2.8 Human serum albumin in complex with compound 2
1HK2 2003-05-16 2.8 HUMAN SERUM ALBUMIN MUTANT R218H COMPLEXED WITH THYROXINE (3,3',5,5'-TETRAIODO-L-THYRONINE)
4LB2 2013-07-24 2.8 X-ray study of human serum albumin complexed with idarubicin
5GIX 2017-07-19 2.8 Human serum albumin-Palmitic acid-Fe(Hn3piT)Cl2
2I30 2006-12-12 2.9 Human serum albumin complexed with myristate and salicylic acid
2XVQ 2010-11-10 2.9 Human serum albumin complexed with dansyl-L-sarcosine
2BXE 2005-09-22 2.95 Human serum albumin complexed with diflunisal
2BXF 2005-09-22 2.95 Human serum albumin complexed with diazepam
5X52 2017-05-17 3.01 Human serum albumin complexed with octanoate and N-acetyl-L-methionine
4N0F 2014-02-05 3.02 Human FcRn complexed with human serum albumin
4HGK 2013-05-08 3.04 Shark IgNAR variable domain
2BXD 2005-09-22 3.05 Human serum albumin complexed with warfarin
2VUF 2008-06-24 3.05 Human serum albumin complexed with fusidic acid
2BXC 2005-09-22 3.1 Human serum albumin complexed with phenylbutazone
1BKE 1999-01-13 3.15 HUMAN SERUM ALBUMIN IN A COMPLEX WITH MYRISTIC ACID AND TRI-IODOBENZOIC ACID
4S1Y 2015-04-29 3.16 X-ray structure of human serum albumin complexed with cisplatin
2BXB 2005-09-22 3.2 Human serum albumin complexed with oxyphenbutazone
1O9X 2003-03-13 3.2 HUMAN SERUM ALBUMIN COMPLEXED WITH TETRADECANOIC ACID (MYRISTIC ACID) AND HEMIN
5IFO 2016-06-01 3.2 X-ray structure of HSA-Myr-KP1019
4K2C 2013-05-01 3.23 HSA Ligand Free
3JQZ 2010-04-21 3.3 Crystal Structure of Human serum albumin complexed with Lidocaine
5FUO 2016-06-29 3.6 Extending the half-life of a Fab fragment through generation of a humanised anti-Human Serum Albumin (HSA) Fv domain: an investigation into the correlation between affinity and serum half-life
4N0U 2014-02-05 3.8 Ternary complex between Neonatal Fc receptor, serum albumin and Fc
6JE7 2019-03-27 3.9 Crystal structure of human serum albumin crystallized by ammonium sulfate

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 SERUM ALBUMIN P02768 ALBU_HUMAN
98.5 SERUM ALBUMIN Q5NVH5 ALBU_PONAB
93.7 Serum albumin A2V9Z4 ALBU_MACFA
93.7 Serum albumin Q28522 ALBU_MACMU