A Crosslinked Cofactor in Lysyl Oxidase: Redox Function for Amino Acid Side Chains

Wang, Sophie Xuefei; Mure, Minae; Medzihradszky, Katalin F.; Burlingame, Alma L.; Brown, Doreen E.; Dooley, David M.; Smith, Alan Jay; Kagan, Herbert M.; Klinman, Judith P.

doi:10.1126/science.273.5278.1078

Cited by 346 publications

(327 citation statements)

References 44 publications

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“…Table I shows that in the native BLO, the copper content was 11 M, and the molar ratio of copper to enzyme was about 1:1 (Table I). The LTQ content of the native BLO was 6 M, meaning that about 50% of the native enzyme was titratable by phenylhydrazine and was catalytically functional, consistent with previous reports (7,12). As shown in Fig.…”

Section: Resultssupporting

confidence: 73%

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The Catalytic Function of Bovine Lysyl Oxidase in the Absence of Copper

Tang

Klinman

2001

Journal of Biological Chemistry

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). By a combination of UV-visible spectroscopy, metal content analysis, and activity measurements, we find that copper-depleted BLO reacts in an irreversible manner with phenylhydrazine, an amine substrate analog, and catalyzes multiple turnovers of the substrate benzylamine. After removal of the majority of enzyme-bound copper, apoBLO exhibits a decrease in the LTQ content, as evidenced by the drop of the 510 -520-nm absorbance, suggesting that the copper may play a structural role in stabilizing the LTQ. The remaining intact LTQ in the apoBLO reacted with phenylhydrazine, both in the presence and absence of the chelator, 10 mM 2,2-dipyridyl. When benzylamine was used as the substrate, the apoBLO turned over at a rate of 50 -60% of the native BLO (after correction for the residual copper and the change of LTQ content). Copper contamination from the assay buffer was ruled out by comparison of enzyme activity using different apoBLO concentrations. These studies demonstrate that the mature form of lysyl oxidase retains many of its functions in the absence of copper.

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Section: Resultssupporting

confidence: 73%

“…Recently, the lysine tyrosyl quinone (LTQ) has been identified as the active site carbonyl cofactor in bovine lysyl oxidase (7). Using the UV-visible spectra of LTQ as a probe, we now report the effects of copper removal on the reactivity of BLO with benzylamine and its analog, phenylhydrazine.…”

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confidence: 99%

The Catalytic Function of Bovine Lysyl Oxidase in the Absence of Copper

Tang

Klinman

2001

Journal of Biological Chemistry

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“…Recently, a quinone cofactor was identified in the active site of LO. It is derived from a crosslinking between a Tyr derivative and a Lys residue in LO and is thus designated lysine tyrosylquinone (18). In WS9-14, these Lys and Tyr residues are conserved, suggesting that WS9-14 may also form such a quinone cofactor by cross-linking at the same site (Fig.…”

Section: Resultsmentioning

confidence: 99%

Regulation of a Novel Gene Encoding a Lysyl Oxidase-related Protein in Cellular Adhesion and Senescence

Saitō

Papaconstantinou

Sato

et al. 1997

Journal of Biological Chemistry

100

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We report here a novel cDNA clone with a predicted protein sequence similar to lysyl oxidase. This fulllength cDNA clone of 3432 base pairs (WS9-14) was isolated from human fibroblasts on the basis of its overexpression in senescent cells. It encodes an 87-kDa polypeptide, whose protein is a member of the scavenger receptor cysteine-rich family, because it contains four scavenger receptor cysteine-rich domains that are found in several secreted or cell surface proteins. The WS9-14 protein has a 48% identity with both lysyl oxidase and lysyl oxidase-like protein at a region corresponding to exons 2-6, implying the existence of a lysyl oxidase gene family. The pattern of WS9-14 gene expression by fibroblasts parallels pro-collagen I-␣1 expression. Its mRNA level is induced by transforming growth factor ␤-1 and indomethacin and inhibited by phorbol ester and retinoic acid. WS9-14 is abundantly expressed in all tumor cell lines examined that attach to culture dishes but not in cell lines that grow in suspension and is also up-regulated in senescent fibroblasts. These results suggest that WS9-14 gene encodes an extracellular protein that may be specifically involved in cell adhesion and senescence.

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“…Analysis of selenium content was performed using a Perkin Elmer 4100 ZL Atomic Absorption Spectrometer. Two separate quinone labeling methods were performed as described (13,14). High and low temperature EPR spectra were recorded on a Bruker ESP-300S spectrometer and a Bruker E580 spectrometer equipped with a ESR-910 liquid helium cryostat (Oxford Instruments), respectively.…”

Section: Methodsmentioning

confidence: 99%

Mechanistic Insights Revealed Through Characterization of a Novel Chromophore in Selenophosphate Synthetase from Escherichia coli

Wolfe

2003

IUBMB Life

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SummaryThe incorporation of selenium into specific proteins and tRNAs requires selenophosphate (SePO 3 ), whose formation is catalyzed by selenophosphate synthetase. In a Mg/ATP-dependent reaction, selenophosphate synthetase catalyzes the phosphorylation of selenide to yield AMP, inorganic phosphate, and SePO 3 . In this report, a previously unrecognized chromophore covalently attached to selenophosphate synthetase is characterized. The UV/Vis spectrum of selenophosphate synthetase has a feature centered at 315 nm that is irreversibly destroyed by alkylation. Moreover, addition of Zn 2 + , which is known to inhibit selenophosphate synthetase, reversibly quenches the 315 nm absorption. Since Zn 2 + is known to bind to Cys17, these data strongly suggest that this residue participates in the 315 nm absorption. Upon incubation with both Mg 2 + and ATP, the l max of the chromophore shifts to 340 nm, and it is shown that the shift requires binding of nucleotide having a hydrolyzable g-phosphoryl group. These data indicate that either the chromophore is directly involved in phosphoryl transfer or indirectly reflects a phosphorylation-dependent conformational change in selenophosphate synthetase. This work provides the first spectroscopic handle on catalytic steps associated with SePO 3 synthesis, which will be used to study the molecular structure of the chromophore and its role in the catalytic mechanism of selenophosphate synthetase. IUBMB Life, 55: 689-693, 2003

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A Crosslinked Cofactor in Lysyl Oxidase: Redox Function for Amino Acid Side Chains

Cited by 346 publications

References 44 publications

The Catalytic Function of Bovine Lysyl Oxidase in the Absence of Copper

The Catalytic Function of Bovine Lysyl Oxidase in the Absence of Copper

Regulation of a Novel Gene Encoding a Lysyl Oxidase-related Protein in Cellular Adhesion and Senescence

Mechanistic Insights Revealed Through Characterization of a Novel Chromophore in Selenophosphate Synthetase from Escherichia coli

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