Primary structure and binding characteristics of locust and human muscle fatty-acid-binding proteins

R. G H J Maatman, M. Degano, H. T B Van Moerkerk, W. J A Van Marrewijk, D. J. Van Der Horst, J. C. Sacchettini, J. H. Veerkamp

Research output: Contribution to journalArticle

Abstract

The conservation between muscle fatty-acid-binding proteins (M-FABP) of Locusta migratoria flight muscle and human skeletal muscle was investigated. The locust M-FABP cDNA (632 bp) was isolated by 5' and 3' rapid amplification of cDNA ends. The identities of the locust and human M-FABP on the cDNA and protein levels were 54% and 42%, respectively. The predicted amino acid sequence of locust M-FABP indicated a molecular mass of 14935 Da and isoelectric point 6.1. The locust M-FABP was expressed in Escherichia coli, purified by (NH4)2SO4 precipitation, anion-exchange and gel-filtration chromatographies and compared with the recombinant human M-FABP with respect to immunological and binding properties. In spite of the high sequence similarity, the proteins did not show immunological cross-reactivity. The binding parameters of locust M-FABP were analyzed with radiolabeled oleic acid by the Lipidex assay and titration microcalorimetry. Both methods revealed a K(d) for oleic acid of 0.5 μM and a binding stoichiometry of 1 mol fatty acid/mol FABP. The ΔH, ΔG and ΔS for oleic acid binding were -146 kJ·mol-1 and -36 J·mol-1 and -369 J·mol-1·K-1 respectively. All the information obtained from binding, fluorescence and displacement studies indicated that locust M-FABP has binding characteristics similar to human MFABP. Finally the recombinant locust M-FABP was crystallized with and without oleic acid. All crystals were trigonal in the P3121 space group. The unit cell dimensions were a = b = 5.89 nm and c = 14.42 nm.

Original languageEnglish
Pages (from-to)801-810
Number of pages10
JournalEuropean Journal of Biochemistry
Volume221
Issue number2
Publication statusPublished - 1994

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Fatty Acid-Binding Proteins
Grasshoppers
Muscle Proteins
Muscle
Oleic Acid
Complementary DNA
Locusta migratoria
Isoelectric Point
Protein Binding
Gel Chromatography
Anions
Molecular mass
Amino Acid Sequence
Chromatography
Titration
Skeletal Muscle
Proteins
Fatty Acids
Stoichiometry
Fluorescence

ASJC Scopus subject areas

  • Biochemistry

Cite this

Maatman, R. G. H. J., Degano, M., Van Moerkerk, H. T. B., Van Marrewijk, W. J. A., Van Der Horst, D. J., Sacchettini, J. C., & Veerkamp, J. H. (1994). Primary structure and binding characteristics of locust and human muscle fatty-acid-binding proteins. European Journal of Biochemistry, 221(2), 801-810.

Primary structure and binding characteristics of locust and human muscle fatty-acid-binding proteins. / Maatman, R. G H J; Degano, M.; Van Moerkerk, H. T B; Van Marrewijk, W. J A; Van Der Horst, D. J.; Sacchettini, J. C.; Veerkamp, J. H.

In: European Journal of Biochemistry, Vol. 221, No. 2, 1994, p. 801-810.

Research output: Contribution to journalArticle

Maatman, RGHJ, Degano, M, Van Moerkerk, HTB, Van Marrewijk, WJA, Van Der Horst, DJ, Sacchettini, JC & Veerkamp, JH 1994, 'Primary structure and binding characteristics of locust and human muscle fatty-acid-binding proteins', European Journal of Biochemistry, vol. 221, no. 2, pp. 801-810.
Maatman RGHJ, Degano M, Van Moerkerk HTB, Van Marrewijk WJA, Van Der Horst DJ, Sacchettini JC et al. Primary structure and binding characteristics of locust and human muscle fatty-acid-binding proteins. European Journal of Biochemistry. 1994;221(2):801-810.
Maatman, R. G H J ; Degano, M. ; Van Moerkerk, H. T B ; Van Marrewijk, W. J A ; Van Der Horst, D. J. ; Sacchettini, J. C. ; Veerkamp, J. H. / Primary structure and binding characteristics of locust and human muscle fatty-acid-binding proteins. In: European Journal of Biochemistry. 1994 ; Vol. 221, No. 2. pp. 801-810.
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AU - Degano, M.

AU - Van Moerkerk, H. T B

AU - Van Marrewijk, W. J A

AU - Van Der Horst, D. J.

AU - Sacchettini, J. C.

AU - Veerkamp, J. H.

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