[논문]Tandem Mass Spectrometric Evidence for the Involvement of a Lysine Basic Side Chain in the Coordination of Zn(II) Ion within a Zinc-bound Lysine Ternary Complex

Yu, Sung-Hyun; Lee, Sun-Young; Chung, Gyu-Sung; Oh, Han-Bin

doi:10.5012/bkcs.2004.25.10.1477

Tandem Mass Spectrometric Evidence for the Involvement of a Lysine Basic Side Chain in the Coordination of Zn(II) Ion within a Zinc-bound Lysine Ternary Complex 원문보기

Bulletin of the Korean chemical society, v.25 no.10, 2004년, pp.1477 - 1483

Yu, Sung-Hyun (Department of Chemistry, Sogang University) , Lee, Sun-Young (Department of Chemistry, Sogang University) , Chung, Gyu-Sung (Department of Chemistry, Konyang University) , Oh, Han-Bin (Department of Chemistry, Sogang University)

Abstract ▼ AI-Helper

We present the tandem mass spectrometry applications carried out to elucidate the coordination structure of Zn(II) bound lysine ternary complexes, $(Zn+Lys+Lys-H)^+$, which is a good model system to represent a simple (metallo)enzyme-substrate complex (ES). In particular, experimental efforts were focused on revealing the involvement of a lysine side chain ${\varepsilon}$-amino group in the coordination of $Zn^{2+}$ divalent ions. MS/MS fragmentation pattern showed that all the oxygen species within a complex fell off in the form of $H_2O$ in contrast to those of other ternary complexes containing amino acids with simple side chains (4-coordinate geometries, Figure 1a), suggesting that the lysine complexes have different coordination structures from the others. The participation of a lysine basic side chain in the coordination of Zn(II) was experimentally evidenced in MS/MS for $N{\varepsilon}$-Acetyl-L-Lys Zn(II) complexes with acetyl protection groups as well as in MS/MS for the ternary complexes with one $NH_3$ loss, $(Zn+Lys+Lys-NH_3-H)^+$. Detailed structures were predicted using ab initio calculations on $(Zn+Lys+Lys-H)^+$ isomers with 4-, 5-, and 6-coordinate structures. A zwitterionic 4-coordinate complex (Figure 7d) and a 5-coordinate structure with distorted bipyramidal geometry (Figure 7b) are found to be most plausible in terms of energy stability and compatibility with the experimental observations, respectively.

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제안 방법

To our surprise, the ions corresponding to abnormal, extensive H2O losses (up to four losses) were observed as major products along with those of a NH3 loss or of Zn(II) bound lysine binary complexes, (64Zn+Lys-H)+. In order to verify whether respective ions contain a Zn atom, the identical experiment was also performed on the second most abundant isotope complexes of (66Zn+Lys+Lys-H)+, and it confirmed our mass spectral interpretation. This unique, abnormal fragmentation behavior was in sharp contrast to that observed in the SORI-CAD on proton-bound lysine dimers of (Lys+Lys +H)+ in which monomeric dissociation into (Lys+H)+ was dominant (data not shown).
It is expected that there may exist a variety of plausible pathways leading to extensive H2O losses, but the scrutiny of all these pathways is, at this current stage of investigations, beyond the scope ofthis report. In order to verify whether these H2O losses took place in sequence or not, daughter complexes with one or two H2O losses were re-isolated and were subjected to the second SORI-CAD. The resulting MS/MS/MS spectrum ofFigure 3 shows that sufficient collision energies given to the daughter ions induced additional H2O losses.

이론/모형

Ionized molecules were externally accumulated for 2 seconds in a hexapole trap before being pulsed into a long single stage quadrupole guide leading to an ion cyclotron resonance (ICR) cell. Trapping of the ions in the ICR cell was made using both gated-trapping and collision gas cooling methods. For MS/MS tandem mass spectrometry studies, the ions of interest were isolated using one broadband SWIFT (Stored Waveform Inverse Fouier Transform) waveform.

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