Functional and conformational characterization of new mutants of heart fatty acid-binding protein.


Abstract

In this study we investigated the possible involvement of several amino acids (not located in the ligand-binding centre) in fatty acid binding and conformational stability of heart fatty acid-binding protein (H-FABP). We prepared recombinant human H-FABP proteins with mutations in the hydrophobic patch (Phe(4), Trp(8) and Phe(64)), portal region (Phe(16)), hinge region (Leu(66), Gly(67)), second portal region (Glu(72)) and at the protein surface (Lys(21)) respectively. Oleic acid-binding affinity and conformational stability of human H-FABP are significantly decreased or completely lost by mutation of Trp(8) or Phe(16). NMR spectra confirmed that these residues are important for the stability of the protein fold. Substitution of Phe(4) or Phe(64) resulted in less stability, but oleic acid-binding affinity was not affected. Mutation of Lys(21) had no effect on either structural integrity or fatty acid-binding affinity. Replacement of Leu(66) or Gly(67) did not affect fatty acid binding, but protein stability was reduced. Finally, mutation of Glu(72) to Ser caused no change of affinity, but NMR spectra and urea-denaturation curves showed the extremely poor stability of this mutant. In conclusion, no relationship was observed between fatty acid-binding affinity and conformational stability. Study holds ProTherm entries: 12966, 12967, 12968, 12969, 12970, 12971, 12972, 12973, 12974, 12975, 12976, 12977 Extra Details: fluorescence; NMR spectroscopy; portal region; sitedirected mutagenesis

Submission Details

ID: LDzqoY684

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:44 p.m.

Version: 1

Publication Details
Zimmerman AW;Rademacher M;Rüterjans H;Lücke C;Veerkamp JH,Biochem. J. (1999) Functional and conformational characterization of new mutants of heart fatty acid-binding protein. PMID:10567233
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
1G5W 2001-03-07 SOLUTION STRUCTURE OF HUMAN HEART-TYPE FATTY ACID BINDING PROTEIN
4TKB 2015-01-28 0.86 The 0.86 angstrom X-ray structure of the human heart fatty acid-binding protein complexed with lauric acid
4TKJ 2015-01-28 0.87 The 0.87 angstrom X-ray structure of the human heart fatty acid-binding protein complexed with palmitic acid
4TJZ 2015-01-28 0.87 The 0.87 angstrom X-ray structure of the human heart fatty acid-binding protein complexed with capric acid
3WVM 2015-01-28 0.88 The 0.88 angstrom X-ray structure of the human heart fatty acid-binding protein complexed with stearic acid
5B28 2017-01-18 0.9 The 0.90A structure of human FABP3 F16V mutant complexed with palmitic acid
4TKH 2015-01-28 0.93 The 0.93 angstrom X-ray structure of the human heart fatty acid-binding protein complexed with myristic acid
5CE4 2016-03-09 0.98 High Resolution X-Ray and Neutron diffraction structure of H-FABP
5B27 2017-01-18 1.02 The 1.02A structure of human FABP3 M20S mutant complexed with palmitic acid
5B29 2017-01-18 1.28 The 1.28A structure of human FABP3 F16V mutant complexed with palmitic acid at room temperature
4WBK 2015-01-28 1.37 The 1.37 angstrom X-ray structure of the human heart fatty acid-binding protein complexed with stearic acid
1HMS 1995-05-08 1.4 1.4 ANGSTROMS STRUCTURAL STUDIES ON HUMAN MUSCLE FATTY ACID BINDING PROTEIN: BINDING INTERACTIONS WITH THREE SATURATED AND UNSATURATED C18 FATTY ACIDS
1HMR 1995-05-08 1.4 1.4 ANGSTROMS STRUCTURAL STUDIES ON HUMAN MUSCLE FATTY ACID BINDING PROTEIN: BINDING INTERACTIONS WITH THREE SATURATED AND UNSATURATED C18 FATTY ACIDS
1HMT 1995-05-08 1.4 1.4 ANGSTROMS STRUCTURAL STUDIES ON HUMAN MUSCLE FATTY ACID BINDING PROTEIN: BINDING INTERACTIONS WITH THREE SATURATED AND UNSATURATED C18 FATTY ACIDS
6AQ1 2018-06-13 1.4 The crystal structure of human FABP3
3WXQ 2014-11-05 1.6 Serial femtosecond X-ray structure of human fatty acid-binding protein type-3 (FABP3) in complex with stearic acid (C18:0) determined using X-ray free-electron laser at SACLA
2HMB 1994-01-31 2.1 THREE-DIMENSIONAL STRUCTURE OF RECOMBINANT HUMAN MUSCLE FATTY ACID-BINDING PROTEIN
3WBG 2013-10-30 2.15 Structure of the human heart fatty acid-binding protein in complex with 1-anilinonaphtalene-8-sulphonic acid
5HZ9 2016-12-14 2.3 human FABP3 in complex with 6-Chloro-2-methyl-4-phenyl-quinoline-3-carboxylic acid
3RSW 2012-05-16 2.6 Crystal Structure of Heart Fatty Acid Binding Protein (FABP3)

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
90.1 Fatty acid-binding protein, heart P07483 FABPH_RAT
91.0 Fatty acid-binding protein, heart Q99P61 FABPH_ICTTR
90.2 Fatty acid-binding protein, heart O02772 FABPH_PIG
100.0 Fatty acid-binding protein, heart P05413 FABPH_HUMAN