STRUCTURE-FUNCTION-RELATIONSHIPS OF MITOCHONDRIAL MONOAMINE-OXIDASE-A AND MONOAMINE-OXIDASE-B CHIMERIC FORMS

被引：39

作者：

GOTTOWIK, J ^{[1
]}

MALHERBE, P ^{[1
]}

LANG, G ^{[1
]}

DAPRADA, M ^{[1
]}

CESURA, AM ^{[1
]}

机构：

[1] F HOFFMANN LA ROCHE & CO LTD, NERVOUS SYST DIS, PRECLIN RES, DIV PHARMA, CH-4002 BASEL, SWITZERLAND

来源：

EUROPEAN JOURNAL OF BIOCHEMISTRY | 1995年 / 230卷 / 03期

关键词：

MONOAMINE OXIDASE; CHIMERIC ENZYME; STRUCTURE FUNCTION RELATIONSHIPS; LAZABEMIDE;

D O I：

10.1111/j.1432-1033.1995.tb20639.x

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Monoamine oxidases (MAO) A and B show a high degree of amino acid similarity. Apart from the NH2-terminus, which contains an ADP-binding consensus sequence, little is known about their structural features or the sequences involved in the binding of substrates. In the present paper, we have studied the structure/function relationships of MAOs by constructing 18 different chimeric forms of MAO, engineered by moving progressively the junction between the NH2-terminus of one MAO form with the COOH-terminus of its isoenzyme. After transient expression in HEK-293 cells, the properties of these chimeric enzymes were investigated using both selective and nonselective substrates and inhibitors. Whereas exchange of the ADP-binding sequence did not modify the catalytic properties of either MAO isoforms, chimeras with increasing length of the NH2-terminus of MAO-A (up to residue 256) showed a marked decrease in affinity towards the MAO-B substrate phenylethylamine and the inhibitor N-(2-aminoethyl)-5-chloro-2-pyridine carboxamide . HCl (lazabemide) when compared to wild-type MAO-B. No major changes were observed in the k(cat) values of these chimeras. From the data obtained, two sequences, i.e. 62-103 and 146-220, appeared of particular importance in constituting the binding site of MAO-B. On the other hand, the catalytic properties and specificity of MAO-A appeared to be relatively insensitive to substitution of both the NH2- (up to position 112) and COOH-termini (from residue 395) of MAO-A with the corresponding MAO-B sequences. However, further modification of the central 283-residue sequence of MAO-A did not appear compatible with enzymic activity. None of the engineered chimeras showed a shift in specificity from one isoform to the other.

引用

页码：934 / 942

页数：9

共 31 条

[1] Bach A.J.W., Lan N.C., Johnson D.L., Abell C.W., Bembenek M.E., Kwan S.W., Seeburg P., Shih J.C., cDNA cloning of human liver monoamine oxidase A and B: molecular basis of differences in enzymic properties, Proc. Natl Acad. Sci. USA, 85, pp. 4934-4938, (1988)
[2] Bertocci B., Miggiano V., Da Prada M., Denbic Z., Lahm H.-W., Malherbe P., Human catechol O‐methyltransferase: cloning and expression of the membrane‐associated form, Proc. Natl Acad. Sci. USA, 88, pp. 1416-1420, (1991)
[3] Birnboim H.C., Doly J., A rapid alkaline extraction procedure for screening recombinant plasmid DNA, Nucleic Acids Res., 7, (1979)
[4] Caron M.G., Chimeras speak again, Nature, 366, (1993)
[5] Cesura A.M., Galva M.D., Imhof R., Kyburz E., Picotti G.B., Da Prada M., [<sup>3</sup>H]Ro 19–6327: a reversible ligand and affinity labelling probe for monoamine oxidase‐A, Eur. J. Pharmacol., 162, pp. 457-465, (1989)
[6] Cesura A.M., Bos M., Galva M.D., Imhof R., Da Prada M., Characterisation of the binding of [<sup>3</sup>H]Ro 41–1049 to the active site of human monoamine oxidase‐A, Mol. Pharmacol., 37, pp. 358-366, (1990)
[7] Cesura A.M., Pletscher A., The new generation of monoamine oxidase inhibitors, Prog. Drug Res., 38, pp. 171-297, (1992)
[8] Da Prada M., Kettler R., Keller H.H., Burkard W.P., Muggli-Manig-lio D., Haefely W.E., Neurochemical profile of moclobemide, a short‐acting and reversible inhibitor of monoamine oxidase type A, J. Pharm. Exp. Ther., 248, pp. 400-414, (1989)
[9] Da Prada M., Kettler R., Keller H.H., Cesura A.M., Richards J.G., Saura Marti J., Muggli-Maniglio D., Wyss P.-C., Kyburz E., Imhof R., From moclobemide to Ro 19–6327 and Ro 41–1049: the development of a new class of reversible, selective MAO‐A and MAO‐B inhibitors, J. Neural Transm. (Suppl.), 9, pp. 45-89, (1990)
[10] Denney R.M., Patel N.T., Fritz R.R., Abell C.W., A monoclonal antibody elicited to human platelet monoamine oxidase: isolation and specificity for human monoamine oxidase B but not A, Mol. Pharmacol., 22, pp. 500-508, (1982)

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