Cleavable Linkers for Porous Silicon‐Based Mass Spectrometry

Meng, Juncai; Averbuj, Claudia; Lewis, Warren G.; Siuzdak, Gary; Finn, M. G.

doi:10.1002/ange.200352803

Cited by 10 publications

(11 citation statements)

References 63 publications

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“…Another method was presented by Meng et al (2004): in order to bind the substrate molecules, the surface of a DIOS chip was derivatized using special linkers, which are stable during the organic synthesis and cleavable during the LDI event. A similar approach was also adopted by Parker et al (2006); here, acrylamide‐labeled peptides were immobilized in a hydrogel surface and the enzymatic activity of a kinase was studied by detecting the phosphorylated peptides (Fig.…”

Section: On‐plate Chemistry and Biochemistrymentioning

confidence: 99%

DNA methyltransferase expression in the mouse germ line during periods of de novo methylation

Lees-Murdock

Shovlin

Gardiner

et al. 2005

Developmental Dynamics

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DNA methyltransferase (DNMT) 3A and DNMT3B are both active de novo DNA methyltransferases required for development, whereas DNMT3L, which has no demonstrable methyltransferase activity, is required for methylation of imprinted genes in the oocyte. We show here that different mechanisms are used to restrict access by these proteins to their targets during germ cell development. Transcriptional control of the Dnmt3l promoter guarantees that message is low or absent except during periods of de novo activity. Use of an alternative promoter at the Dnmt3a locus produces the shorter Dnmt3a2 transcript in the germ line and postimplantation embryo only, whereas alternative splicing of the Dnmt3b transcript ensures that Dnmt3b1 is absent in the male prospermatogonia. Control of subcellular protein localization is a common theme for DNMT3A and DNMT3B, as proteins were seen in the nucleus only when methylation was occurring. These mechanisms converge to ensure that the only time that functional products from each locus are present in the germ cell nuclei is around embryonic day 17.5 in males and after birth in the growing oocytes in females. Developmental Dynamics 232:992-1002, 2005.

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Section: On‐plate Chemistry and Biochemistrymentioning

confidence: 99%

DNA methyltransferase expression in the mouse germ line during periods of de novo methylation

Lees-Murdock

Shovlin

Gardiner

et al. 2005

Developmental Dynamics

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“…However, Finn, Siuzdak, and co-workers have reported a related approach that employs cleavable linkers and may offer another route to high-throughput applications. 18,19 Additionally, the RapidFire method allows high-throughput analysis using mass spectrometry but requires a solid phase extraction or chromatographic sample preparation of the analyte before mass analysis. (4) With SAMDI, the array plates (384 spots) need only be rinsed and treated with matrix after analyte immobilization.…”

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confidence: 99%

High-Throughput Screening of Small Molecule Libraries using SAMDI Mass Spectrometry

et al. 2011

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High-throughput screening is a common strategy used to identify compounds that modulate biochemical activities, but many approaches depend on cumbersome fluorescent reporters or antibodies and often produce false-positive hits. The development of “label-free” assays addresses many of these limitations, but current approaches still lack the throughput needed for applications in drug discovery. This paper describes a high-throughput, label-free assay that combines self-assembled monolayers with mass spectrometry, in a technique called SAMDI, as a tool for screening libraries of 100 000 compounds in one day. This method is fast, has high discrimination, and is amenable to a broad range of chemical and biological applications.

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“…7−9 Based on a similar fragmentation behavior observed for furan−maleimide adducts, this transition likely occurs via a retro DA (rDA) mechanism (Scheme 3). 43 Given this apparent labile nature of fulvene−maleimide adducts in the gas phase, we sought to determine if a similar fragmentation pattern would occur for conjugates without the presence of a photolabile group that might enable the development of a new class of mass tag reagents for biomolecule detection.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

“…At present, we could only find one other example where MS-induced rDA fragmentation has been exploited for the generation of mass tags, in which furan− maleimide adducts were utilized for monitoring enzymecatalyzed reactions of immobilized substrates on silicon chips using a MALDI-TOF instrument. 43 Despite their efficient fragmentation, the subdued reaction kinetics of furan− maleimide cycloadditions and reduced stability under physiological conditions limit their utility outside of organic synthesis. In contrast, we have shown that the fulvene−maleimide DA reaction can be used to rapidly generate protein bioconjugates in aqueous buffer at low temperatures and has facile fragmentation properties that enable mass tag detection in both MALDI and ESI experiments.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

“…In contrast, we have shown that the fulvene−maleimide DA reaction can be used to rapidly generate protein bioconjugates in aqueous buffer at low temperatures and has facile fragmentation properties that enable mass tag detection in both MALDI and ESI experiments. 8,43 Here, we further utilize these favorable properties to demonstrate the detection of protein−peptide conjugates directly from complex biological samples.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

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Retro Diels–Alder Fragmentation of Fulvene–Maleimide Bioconjugates for Mass Spectrometric Detection of Biomolecules

et al. 2021

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Diels−Alder chemistry is a well-explored avenue for the synthesis of bioactive materials; however, its potential applications have recently expanded following the development of reactions that can be performed in buffered aqueous environments at low temperatures, including fulvene−maleimide [4 + 2] cycloadditions. In this study, we synthesized two novel amine-reactive fulvene linkers to demonstrate the application of this chemistry for generating mass spectrometry-cleavable labels ("mass tags"), which can be used for the labeling and detection of proteins. Successful conjugation of these linkers to maleimidelabeled peptides was observed at low temperatures in phosphatebuffered saline, allowing the non-destructive modification of proteins with such mass tags. The labile nature of fulvene−maleimide adducts in the gas phase also makes them suitable for both matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization (ESI) mass spectrometric analysis. Unlike previous examples of MALDI mass tags, we show that fulvene−maleimide cycloaddition adducts fragment predictably upon gas-phase activation without the need for bulky photocleavable groups. Further exploration of this chemistry could therefore lead to new approaches for mass spectrometry-based bioassays.

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Cleavable Linkers for Porous Silicon‐Based Mass Spectrometry

Abstract: Dippen – waschen – analysieren: Moleküle können durch Diels‐Alder‐Reaktionen an maßgeschneiderte poröse Silicium‐Wafer geknüpft und mit dem Laserpuls eines MALDI‐Massenspektrometers positionscodiert effizient abgelöst und detektiert werden (siehe Schema).

Cited by 10 publications

References 63 publications

DNA methyltransferase expression in the mouse germ line during periods of de novo methylation

DNA methyltransferase expression in the mouse germ line during periods of de novo methylation

High-Throughput Screening of Small Molecule Libraries using SAMDI Mass Spectrometry

Retro Diels–Alder Fragmentation of Fulvene–Maleimide Bioconjugates for Mass Spectrometric Detection of Biomolecules

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