Fatty acid binding and conformational stability of mutants of human muscle fatty acid-binding protein.


Abstract

Human muscle fatty acid-binding protein (M-FABP) is a 15 kDa cytosolic protein which may be involved in fatty acid transfer and modulation of non-esterified fatty acid concentration in heart, skeletal muscle, kidney and many other tissues. Crystallographic studies have suggested the importance of the amino acids Thr-40, Arg-106, Arg-126 and Tyr-128 for the hydrogen bonding network of the fatty acid carboxylate group. Two phenylalanines at 16 and 57 are positioned to interact with the acyl chain of the fatty acid. We prepared 13 mutant proteins by site-directed mutagenesis and tested them for fatty acid binding and stability. Substitution of amino acids Phe-16, Arg-106 or Arg-126 created proteins which showed a large decrease in or complete loss of oleic acid binding. Substitution of Phe-57 by Ser or Val and of Tyr-128 by Phe had no great effect. The stability of the mutant proteins was tested by denaturation studies on the basis of fatty acid binding or tryptophan fluorescence and compared with that of the wild-type M-FABP. There was no direct relationship between fatty acid-binding activity and stability. Less stable mutants (F57S and Y128F) did not show a marked change in fatty acid-binding activity. Substitution of Arg-126 by Gln or Arg-106 by Thr eliminated binding activity, but the former mutant protein showed wild-type stability, in contrast to the latter. The results are in agreement with crystallographic data. Study holds ProTherm entries: 7258, 7259, 7260, 7261, 7262, 7263, 7264, 7265, 7266, 7267, 7268, 7269, 7270 Extra Details:

Submission Details

ID: RvYhZxg64

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:33 p.m.

Version: 1

Publication Details
Prinsen CF;Veerkamp JH,Biochem. J. (1996) Fatty acid binding and conformational stability of mutants of human muscle fatty acid-binding protein. PMID:8660291
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