Mass Spectrometry-Based Disulfide Mapping of Lysyl Oxidase-like 2

Meier, Alex; Go, Eden P.; Moon, Hee-Jung; Desaire, Heather; Mure, Minae

doi:10.3390/ijms23115879

Cited by 7 publications

(11 citation statements)

References 35 publications

(53 reference statements)

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“…Since Zn 2+ cannot support the activation/oxidation of Tyr689, the unproductive conformation of Lys653 seems to be thermodynamically favored during protein crystallization. However, our solution study of the catalytically–competent LOXL2 strongly indicates that β8 and the Lys653 sidechain are not fixed in the unproductive conformation seen in the precursor structure: instead, they can accommodate the productive conformation of Lys653 to support the LTQ cofactor biogenesis in the presence of Cu 2+ under aerobic conditions [ 19 , 24 ].…”

Section: Resultsmentioning

confidence: 99%

“…LOXL2 contains the lysine tyrosylquinone (LTQ) cofactor that is post–translationally derived from Lys653 and Tyr689 in the C–terminal catalytic domain [ 18 , 19 ]. Based on the mechanism of the topaquinone (TPQ) cofactor biogenesis in copper amine oxidases (CAOs), it has been proposed that a 1,4–addition (Michael addition) of the ϵ–amino group of Lys653 to dopaquinone (DPQ) intermediate derived from Tyr689, followed by O 2 –oxidation yields LTQ [ 20 , 21 ].…”

Section: Introductionmentioning

confidence: 99%

“…These results suggest that the overall structures of the precursor and the mature LOXL2 are very similar. Subsequently, we conducted mass spectrometry–based disulfide mapping of the precursor LOXL2 and the mature LOXL2 that was inhibited by 2–hydrazinopyridine (2HP) and demonstrated that the cysteine–pairing pattern of disulfide bonds is unaltered before and after LTQ biogenesis [ 19 ]. These results support our hypothesis that a substantial conformational rearrangement is not required for LTQ biogenesis.…”

Section: Introductionmentioning

confidence: 99%

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A 3D–Predicted Structure of the Amine Oxidase Domain of Lysyl Oxidase–Like 2

Meier

Kuczera

Mure

2022

IJMS

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Lysyl oxidase–like 2 (LOXL2) has been recognized as an attractive drug target for anti–fibrotic and anti–tumor therapies. However, the structure–based drug design of LOXL2 has been very challenging due to the lack of structural information of the catalytically–competent LOXL2. In this study; we generated a 3D–predicted structure of the C–terminal amine oxidase domain of LOXL2 containing the lysine tyrosylquinone (LTQ) cofactor from the 2.4Å crystal structure of the Zn2+–bound precursor (lacking LTQ; PDB:5ZE3); this was achieved by molecular modeling and molecular dynamics simulation based on our solution studies of a mature LOXL2 that is inhibited by 2–hydrazinopyridine. The overall structures of the 3D–modeled mature LOXL2 and the Zn2+–bound precursor are very similar (RMSD = 1.070Å), and disulfide bonds are conserved. The major difference of the mature and the precursor LOXL2 is the secondary structure of the pentapeptide (His652–Lys653–Ala654–Ser655–Phe656) containing Lys653 (the precursor residue of the LTQ cofactor). We anticipate that this peptide is flexible in solution to accommodate the conformation that enables the LTQ cofactor formation as opposed to the β–sheet observed in 5ZE3. We discuss the active site environment surrounding LTQ and Cu2+ of the 3D–predicted structure.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A 3D–Predicted Structure of the Amine Oxidase Domain of Lysyl Oxidase–Like 2

Meier

Kuczera

Mure

2022

IJMS

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“…We have recently identified 2HP-modified LTQ, LTQ-2HP (Fig. 1) in 2HPinhibited LOXL2 by mass spectrometry-based peptide mapping (Meier, Go et al, 2022). Since there is no structural information of a catalytically competent form of LOXL2, we conducted comparative spectroscopic studies of 2HP-inhibited LOXL2 and the corresponding model compound in solution, in order to understand the spatial arrangement of the LTQ cofactor and the active site Cu II .…”

Section: Chemical Contextmentioning

confidence: 99%

“…After the tautomerization of the hydrazone to the azo form, LTQ-2HP ligates to the active site Cu 2+ . The 2HP-modified LTQ (LTQ-2HP) containing the peptide was detected by mass spectrometry (Meier, Go, et al, 2022). Based on the close resemblances of UV-vis and resonance Raman spectra of 2HP-inhibited LOXL2 and the model compound 2, we hypothesize that LTQ-2HP serves as a tridentate ligand to the active site Cu II in LOXL2 .…”

Section: Research Communicationsmentioning

confidence: 99%

Crystal structures of di-μ-chlorido-bis({(E)-5-(ethylamino)-4-methyl-2-[(pyridin-2-yl)diazenyl]phenolato}copper(II)) and chloridobis(1,10-phenanthroline)copper(II) chloride tetrahydrate

Meier

Mian

et al. 2023

Acta Cryst E

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The dark-red title complex crystallized from an equimolar methanol solution of (E)-5-(ethylamino)-4-methyl-2-[(pyridin-2-yl)diazenyl]phenol and CuCl2(phen) (phen = 1,10-phenanthroline) as a centrosymmetric dimer, [CuCl(C14H15N4O)]2. The Cu atoms are bridged by two Cl ligands and have a slightly distorted square-pyramidal coordination, where two N atoms from the azo and the pyridine moieties, a phenolic O and a Cl atom comprise the base and the other Cl occupies the apex position. The apical Cu—Cl bond, 2.6192 (4) Å, is longer than the basal one, 2.2985 (3) Å, due to Jahn–Teller distortion. The dimers are associated via weak intermolecular hydrogen bonds and π–π stacking interactions between phenyl and pyridine rings. A monomeric by-product of the same reaction, [CuCl(phen)2]Cl·4H2O, has a trigonal–bipyramidal coordination of Cu with equatorial Cl ligand, and extensive outer-sphere disorder. In the structure of 4, the packing of cations leaves continuous channels containing disordered Cl− anions and solvent molecules. The identity of the solvent (water or a water/methanol mixture) was not certain. The disordered anion/solvent regions comprise 28% of the unit-cell volume. The disorder was approximated by five partly occupied positions of the Cl− anion and ten positions of O atoms with a total occupancy of 3, giving a total of 48 electrons per asymmetric unit, in agreement with the integral electron density of 47.8 electrons in the disordered region, as was estimated using the BYPASS-type solvent-masking program [van der Sluis & Spek (1990). Acta Cryst. A46, 194–201].

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Conformation Changes Associated with the Formation and Activation of Lysine Tyrosylquinone of LOXL2 Studied by Metadynamics Simulation

Wang,

Lin,

et al. 2024

Advcd Theory and Sims

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Lysyl oxidase‐like 2 (LOXL2) is a Cu2+‐dependent amine oxidase that catalyzes the oxidative deamination of lysine residues of elastin and collagen to generate aldehyde groups. In the active state of LOXL2, its conserved catalytic domain has a lysine tyrosylquinone (LTQ) cofactor and a copper ion. LTQ is post‐translationally derived from Lys653 and Tyr689 of LOXL2. In this study, well‐tempered metadynamics simulations are used to investigate the conformation modulation from the inactive state to active state and obtain the free energy landscape. Interestingly, the simulations show that LTQ precursor residues cannot get close in Zn2+−bound LOXL2, whereas the copper ion binding triggers a conformation change to form an optional structure for LTQ biosynthesis. It is found that a prominent loop, consisting of residues 654–658, hinders the formation of LTQ in Zn2+−LOXL2 by preventing Lys653 from moving into active site of LOXL2, whereas this loop is relatively flexible in Cu2+−LOXL2. After the formation of LTQ, it needs to be activated by adjusting its position to be exposed to the solvent, thus allowing it to react with its substrate proteins. The results indicate that this process involves an intermediate state and that the activation of LTQ overcomes an energy barrier of 5.9 kcal mol−1.

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Mass Spectrometry-Based Disulfide Mapping of Lysyl Oxidase-like 2

Cited by 7 publications

References 35 publications

A 3D–Predicted Structure of the Amine Oxidase Domain of Lysyl Oxidase–Like 2

A 3D–Predicted Structure of the Amine Oxidase Domain of Lysyl Oxidase–Like 2

Crystal structures of di-μ-chlorido-bis({(E)-5-(ethylamino)-4-methyl-2-[(pyridin-2-yl)diazenyl]phenolato}copper(II)) and chloridobis(1,10-phenanthroline)copper(II) chloride tetrahydrate

Conformation Changes Associated with the Formation and Activation of Lysine Tyrosylquinone of LOXL2 Studied by Metadynamics Simulation

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