Expression and Purification of Enzymatically Active Forms of the Human Lysyl Oxidase-like Protein 4

Moon Suk Kim, Sung Su Kim, Sang Taek Jung, Jung Young Park, Han Wook Yoo, Jesang Ko, Katalin Csiszar, Sang Yun Choi, Youngho Kim

    Research output: Contribution to journalArticlepeer-review

    51 Citations (Scopus)

    Abstract

    The lysyl oxidase-like protein 4 (LOXL4) is the latest member of the emerging family of lysyl oxidases, several of which were shown to function as copper-dependent amine oxidases catalyzing lysine-derived cross-links in extracellular matrix proteins. LOXL4 contains four scavenger receptor cysteine-rich domains in addition to the characteristic domains of the LOX family, including the copper-binding domain, the cytokine receptor-like domain, and the residues of the lysyl-tyrosyl quinone cofactor. In an effort to assess its amine oxidase activity, we expressed LOXL4 as recombinant forms attached with hexa-histidine residues at the carboxyl terminus by using an Escherichia coli expression system. The recombinant proteins were purified with nickel-chelating affinity chromatography and converted into enzymatically active forms by stepwise dialysis. The purified LOXL4 proteins showed β-aminopropionitrile-inhibitable activity of 0.022-0.032 units/mg toward a nonpeptidyl substrate, benzylamine. These results indicate that LOXL4, with the four scavenger receptor cysteine rich domains, may also function as an active amine oxidase. Availability of the pure and active forms of LOXL4 will be significantly helpful in functional studies related to substrate specificity and crystal structure of this amine oxidase, which should provide significant insights into functional differences within the LOX family members.

    Original languageEnglish
    Pages (from-to)52071-52074
    Number of pages4
    JournalJournal of Biological Chemistry
    Volume278
    Issue number52
    DOIs
    Publication statusPublished - 2003 Dec 26

    ASJC Scopus subject areas

    • Biochemistry
    • Molecular Biology
    • Cell Biology

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