Identification of Inorganic Elements in Proteins in Human Serum and in DNA Fragments by Size Exclusion Chromatography and Inductively Coupled Plasma Mass Spectrometry with a Magnetic Sector Mass Spectrometer

Wang, Jin; Houk, R. S.; Dreessen, Dawn; Wiederin, Daniel R.

doi:10.1021/ja9735970

Cited by 38 publications

(21 citation statements)

References 28 publications

(16 reference statements)

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“…1 and 2). Thus, the nebulizer and desolvation system do not broaden the chromatographic peaks greatly, at least for the relatively low-resolution separations obtained by SEC, in agreement with previous work (14,15).…”

Section: Effect Of Ionic Strength and Selection Of Mobile Phasesupporting

confidence: 90%

“…For the ions studied in the present work, either low or medium resolution was adequate (14,15). The accelerating voltage was nominally 8 kV.…”

Section: Methodsmentioning

confidence: 98%

“…The column is suitable for separating proteins with molecular weight (MW) from 10,000 to 500,000 Da. This column is larger than those used previously (14,15) and is capable of better separations, but it requires larger samples and has a longer void time. Chromatograms for Ca and Mg showed only small molecules containing these elements, so the shorter, faster column described previously was adequate.…”

Section: Methodsmentioning

confidence: 98%

“…A different column (GPC 300 at 0.1 ml/min) (14,15) was adequate for the measurement of Ca and Mg. Proteins elute much faster from this smaller column than for the one used for the results shown above. Figure 8 shows chromatograms for Ca and Mg in liver extract.…”

Section: Fig 4 Chromatograms Formentioning

confidence: 99%

“…Two previous papers from our group describe the use of SEC-ICP-MS with a magnetic sector instrument for analysis of serum and DNA solutions (14,15). Many unusual elements can be measured with very good signal-to-noise ratio without preconcentration procedures, despite the dilution that accompanies the chromatographic separation.…”

mentioning

confidence: 99%

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Measurement of Trace Elements in Proteins Extracted from Liver by Size Exclusion Chromatography-Inductively Coupled Plasma-Mass Spectrometry with a Magnetic Sector Mass Spectrometer

Wang

Dreessen

Wiederin

et al. 2001

Analytical Biochemistry

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Section: Effect Of Ionic Strength and Selection Of Mobile Phasesupporting

confidence: 90%

“…For the ions studied in the present work, either low or medium resolution was adequate (14,15). The accelerating voltage was nominally 8 kV.…”

Section: Methodsmentioning

confidence: 98%

Section: Methodsmentioning

confidence: 98%

Section: Fig 4 Chromatograms Formentioning

confidence: 99%

mentioning

confidence: 99%

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Measurement of Trace Elements in Proteins Extracted from Liver by Size Exclusion Chromatography-Inductively Coupled Plasma-Mass Spectrometry with a Magnetic Sector Mass Spectrometer

Wang

Dreessen

Wiederin

et al. 2001

Analytical Biochemistry

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Environmental and Clinical Applications of Inductively Coupled Plasma Spectrometry

Vonderheide¹,

Sadi²,

Sutton³

et al. 2006

Inductively Coupled Plasma Spectrometry and Its Applications

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Site‐Selective Photocleavage of Proteins by Uranyl Ions

Duff¹,

Kumar²

2005

Angewandte Chemie

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Dedicated to Professor J. C. Scaiano on the occasion of his 60th birthdayThe uranyl ion (UO 2 2+ ) has a high affinity for the phosphate groups of DNA/RNA, [1] and this affinity was exploited to photocleave these biopolymers at the uranyl-binding sites. [2][3][4][5] Earlier success in our laboratory with site-specific photocleavage of proteins by organic molecules [6] and metal complexes [7] led to the idea that photoactive [8] metal-oxo species such as UO 2 2+ should be capable of cleaving proteins with high selectivity. However, even though there is ample evidence to suggest that UO 2 2+ binds to proteins, [9, 10] it is not known if UO 2 2+ can photocleave the peptide backbone. It is also not known if binding of UO 2 2+ at specific sites would result in high selectivity for the protein cleavage. In addition, the ease with which UO 2 2+ can be directed to metal-binding sites on proteins and the activation of the cation at visible wavelengths indicated that this would be a promising approach for footprinting metalbinding sites. In this report, high-affinity binding of UO 2 2+ to proteins and their site-selective photocleavage, with visible light, is demonstrated for the first time.Serum albumin (SA) and transferrin are known to bind UO 2 2+ , [11,12] and these proteins therefore provided good initial targets to test the above approach. Quantitative data on the binding of UO 2 2+ to bovine SA (BSA), however, were lacking; these were obtained from calorimetric and spectroscopic studies. Isothermal titration calorimetry of the addition of a solution of UO 2 2+ (0.5 mm) to BSA (0.1 mm ; see Supporting Information) indicated exothermic binding (Figure 1). Best fits to the data required a two-site binding model [13] and indicated a high-affinity site (binding constant (K b ) of (1.6 AE 0.7) 10 7 m À1 ) with moderate exothermicity (binding enthalpy (DH) of (À4.7 AE 0.7) kcal mol À1 ) and a low-affinity site (K b = 2.8 AE 2.5 10 5 m À1 ) with weak exothermicity (DH = À(1.2AE0.8) kcal mol À1 ). Binding of UO 2 2+ to BSA was also supported by absorption, circular dichroism (CD), and fluorescence studies.The CD spectra, for example, of a mixture of BSA (1 mm) and UO 2 2+ in increasing concentrations (0-0.8 mm) showed the growth of new bands at 440 and 510 nm (Figure 2). These evidences established the binding of UO 2 2+ to BSA. Photocleavage of BSA by UO 2 2+ was achieved by irradiating a mixture of BSA and UO 2 2+ at 420 nm (Figure 3). Three distinct product bands at 35, 33 and 30 kDa were obtained in yields (11 AE 0.5), (4 AE 0.6), and (11 AE 1) % (lane 5), respectively. No photocleavage was noted without UO 2 2+ (lane 3) or without light (lane 4). Photocleavage was observed at wavelengths up to 460 nm, although the yields were lower (data not shown).To demonstrate the general nature of the photocleavage, other proteins such as human SA (HSA), porcine SA (PSA), glucose oxidase, and transferrin were also tested. These are cleaved with high selectivity (see Supporting Information). As negative controls, we also tested the ...

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Identification of Inorganic Elements in Proteins in Human Serum and in DNA Fragments by Size Exclusion Chromatography and Inductively Coupled Plasma Mass Spectrometry with a Magnetic Sector Mass Spectrometer

Cited by 38 publications

References 28 publications

Measurement of Trace Elements in Proteins Extracted from Liver by Size Exclusion Chromatography-Inductively Coupled Plasma-Mass Spectrometry with a Magnetic Sector Mass Spectrometer

Measurement of Trace Elements in Proteins Extracted from Liver by Size Exclusion Chromatography-Inductively Coupled Plasma-Mass Spectrometry with a Magnetic Sector Mass Spectrometer

Environmental and Clinical Applications of Inductively Coupled Plasma Spectrometry

Site‐Selective Photocleavage of Proteins by Uranyl Ions

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