Chemical derivatization assisted

42 methods · 1999–2026

Chemical derivatization assisted: Covalent modification of the peptide, usually at a terminus, to change how it fragments: selective cleavage, brighter low-mass ions, or a strongly directed ion series.

Covalent modification of the peptide, usually at a terminus, to change how it fragments: selective cleavage, brighter low-mass ions, or a strongly directed ion series.

The earliest of its 42 methods is PSD MALDI high-sensitivity de novo sequencing (1999); 41 more have followed.

Methods 42
Papers describing them 49
Authors 208
Active 1999-06-22 to 2026-06-30
Kinds adjacent (22), algorithm (18), downstream-application (2)
Acquisition DDA (39)

Methods (42)

Oldest first, by the paper that describes each one.

  • PSD MALDI high-sensitivity de novo sequencing (1999): High-sensitivity postsource decay MALDI method for de novo peptide sequencing with N-terminal sulfonation.
  • Charge derivatization MALDI-PSD de novo sequencing (1999): Charge-derivatizes a proteolytic digest before MALDI post-source decay so the fragment series reads cleanly, evaluated on a phosphoenolpyruvate carboxykinase of known structure and then used to sequence an unknown variant from Anaerobiospirillum succiniciproducens.
  • C-terminal ANSA derivatization for negative-ion de novo sequencing (2000): Derivatizes the peptide C-terminus with 4-aminonaphthalenesulfonic acid so high-energy negative-ion CID gives only C-terminal fragment ions, which can be read de novo.
  • Lysine derivatization PSD de novo sequencing (2000): Derivatization workflow for lysine-terminated tryptic peptides to facilitate PSD MALDI de novo sequencing.
  • ZipTip SPITC derivatization for MALDI-PSD de novo sequencing (2004): Sulfonates tryptic peptides with 4-sulfophenyl isothiocyanate on C18 microZipTips so MALDI post-source decay spectra show only y ions, which are then read de novo to identify gel spots.
  • N-terminal SPITC sulfonation for LC-MS/MS de novo sequencing (2004): Selective N-terminal sulfonation of tryptic peptides with 4-sulfophenyl isothiocyanate, adapted to online capillary LC-MS/MS so the spectra are dominated by y ions and read de novo.
  • N-sulfonated peptide MALDI TOF/TOF de novo sequencing (2004): N-sulfonation before MALDI TOF/TOF directs fragmentation into a single dominant series, so a peptide mass fingerprint that does not identify a protein on its own can be followed by a readable de novo spectrum.
  • CHASE (2005): Charge Assisted Sequencing Engine: derivatizes tryptic peptides at the N-terminus with a negatively charged reagent, reads the resulting PSD/TOF-TOF spectra de novo and passes sequences to BLAST.
  • Peroxycarbamate N-terminal labelling (2006): N-terminal t-butyl peroxycarbamate derivatives lose the N-terminal side chain in CID, identifying that residue and labelling the b-ion series to aid de novo sequencing.
  • IRMPD of N-terminal sulfonated peptides (2006): Infrared multiphoton dissociation of N-terminal sulfonated peptides in a quadrupole ion trap, which both dissociates them efficiently and carries the y-ion series all the way down to y1, so the C-terminal residue is not lost.
  • Charged tag with C-terminal digestion (2007): Couples a fixed-charge N-terminal derivatization with a C-terminal digestion of the derivatized peptide, so MALDI spectra carry one interpretable series and the ladder can be read directly.
  • N-terminal fragment isolation de novo sequencing (2007): Isolates protein N-terminal peptides specifically, through reduction, S-alkylation, guanidination and a cleavable biotin coupling, so several proteins can be N-terminally sequenced in parallel at low picomole level.
  • AnchorChip on-target SPITC derivatization (2008): An on-target AnchorChip clean-up for 4-sulfophenyl isothiocyanate derivatized tryptic peptides that replaces reverse-phase desalting before MALDI TOF-TOF de novo sequencing; applied to proteins from fish liver gels.
  • Disulfide oxidation for de novo MALDI sequencing (2008): Shows that performic acid oxidation of the C-terminal cystine ring in frog skin peptides gives more b and y ions than reduction and alkylation, making MALDI de novo sequencing easier.
  • Lys Tag (2008): A single robust lysine modification before MALDI TOF/TOF that improves de novo readability without a multi-step workup, aimed at identifying proteins confidently from biological samples.
  • TMPP arginine-peptide de novo sequencing (2008): Extends TMPP-Ac-OSu tagging to arginine-containing peptides, whose fragmentation the basic side chain otherwise complicates, simplifying the pattern enough to read a sequence.
  • TMPP N-terminal peptide de novo sequencing (2008): Tags peptide N-termini with TMPP-Ac-OSu after metalloendopeptidase digestion and isolates them, giving MALDI spectra of N-terminal peptides clean enough to sequence de novo.
  • N-terminal peptide isolation with CAF de novo sequencing (2008): Isolates protein N-terminal peptides on DITC resin, whether or not the N-terminus is blocked, then sequences them de novo by MALDI-TOF using chemically assisted fragmentation or MS/MS.
  • Derivatization plus ETcaD de novo (2009): N-terminally derivatized, multiply charged peptides are charge-reduced by electron transfer and then collisionally activated to give almost only z-ions for de novo sequencing.
  • Acetylation-enhanced b-ion MALDI sequencing (2010): Acetylates tryptic peptides in methanol to raise b-ion intensity in MALDI TOF/TOF spectra, making the sequence ladders easier to read and validate. Applied to proteins of the unsequenced Antarctic bacterium Pseudomonas syringae.
  • N-terminal tagging for de novo of short peptides (2010): Modifies the N-terminus of short frog tryptophyllin peptides to stop b-ion head-to-tail cyclization and scrambling, giving spectra that can be sequenced de novo by ESI and MALDI MS/MS.
  • Phosphorylation derivatization for de novo sequencing (2011): Phosphorylates peptide N-termini so that UPLC-MS/MS spectra show enhanced a1 and b-ion series and separate glutamine from lysine, easing de novo peptide sequencing.
  • High proton affinity N-terminal derivatization (2011): Guanidino and amidino N-terminal derivatization reagents that strengthen b-ion series under low-energy CID for easier de novo sequencing.
  • Oxazolone C-terminal derivatization (2011): Selective C-terminal derivatization, which is hard because C-terminal and side-chain carboxyls behave alike, improved in efficiency and given a bromine signature so the C-terminal fragment can be picked out and sequenced.
  • Br-signature N-terminal derivatization (2012): A MALDI signal-enhancing N-terminal derivatization carrying a bromine isotope signature, so the tagged N-terminal peptide is unmistakable in the spectrum. Used to read N-terminal processing such as methionine or signal-peptide removal, and to annotate uncharacterized proteins.
  • 193 nm UVPD of imidazolinylated Lys-N peptides (2012): Imidazolinylation of Lys-N peptides followed by 193 nm ultraviolet photodissociation, which yields a SINGLE product ion series instead of the overlapping series that defeat most de novo strategies.
  • Lactobacillus brevis starvation proteome by CAF-/CAF+ de novo (2013): Uses negative- and positive-mode chemically activated fragmentation of derivatized peptides to de novo sequence proteins of Lactobacillus brevis, revealing 36 proteins changed after long-term starvation.
  • CNBr ring-opening of cyclic OBOC peptides (2013): Places methionine in the macrocycle and linker so cyanogen bromide linearises and releases cyclic library peptides, which are then sequenced manually and with de novo software.
  • 351 nm UVPD de novo sequencing (2013): N-terminal chromophore derivatisation makes a peptide absorb at 351 nm, so selective UV photodissociation cleaves it into a simplified y-ion series instead of the mixed ladders CID gives, deliberately to make the spectra amenable to de novo sequencing.
  • 14N/15N Aba charge derivatization (2013): Derivatizes peptide N-termini with 14N- or 15N-labeled 4-amidinobenzoic acid so b-ions show as doublets and improved CID fragmentation simplifies de novo sequencing.
  • Br-TEMPO FRIPS de novo sequencing (2015): Adds bromine to the free-radical-initiated peptide sequencing reagent so N-terminal fragments carry a doublet signature that eases de novo reading.
  • DUST and SPITC CAF MALDI de novo sequencing (Zagreb) (2016): SPITC derivatization of tryptic peptides, done at acidic pH to avoid side reactions, with chemically activated fragmentation on MALDI and the DUST de novo tool tuned to SPITC chemistry.
  • Combinatorial acetylation/guanidinylation labeling (2017): Combined acetylation and guanidinylation of peptides gives equivalent b- and y-ion representation in ion-trap MS/MS, allowing accurate de novo annotation.
  • Carbamylation-assisted de novo sequencing (2018): Studies carbamylation and dual modification of peptides to enrich b- and y-ion series, including b1, for easier de novo sequencing by Q-TOF.
  • Scorpion venom chemical-modification de novo (2019): Chemical modification-assisted de novo sequencing of antimicrobial peptides in scorpion venom.
  • N-terminal imine derivatization (2020): Studies the CID fragmentation of N-terminal peptide imines as a derivatization that enhances de novo peptide sequencing.
  • FBSA N-terminal derivatization (2023): N-terminal derivatization with 4-formyl-benzenesulfonic acid under charge-reduction conditions, so overlapping positive- and negative-mode spectra pinpoint the b-ion series for de novo sequencing.
  • Tetrazine-cyclized peptide UV ring-opening (2023): Designs cyclic peptides on solid phase around a tetrazine so that UV light opens the ring and releases them, turning a cyclic sequence – which tandem MS deconvolutes badly, and which most approved peptide drugs are – into a linear one that sequences de novo.
  • Bis-cysteine macrocycle chemical linearization (2023): Cleaves synthetic bis-Cys macrocyclic peptides at a defined backbone position by Edman chemistry so that mixtures from macrocycle libraries can be sequenced de novo by nLC-MS/MS.
  • Thioether macrocycle oxidative linearization (2024): Oxidising the thioether of a thioether-closed macrocyclic peptide makes collision-induced dissociation open the ring at one site, so the linearised peptide can be sequenced de novo; demonstrated on a randomised macrocycle library.
  • Me-Cou N-terminal derivatization de novo sequencing (2025): N-terminal derivatization with N-succinimidyl 7-methoxycoumarin-3-carboxylate (Me-Cou), which boosts low-mass b-ion yield so de novo sequencing can read the N-terminal residues, applied to untargeted peptidomics on LC-TIMS-qTOF/MS.
  • PSMtags (2025): Small-molecule peptide conjugates found by systematic screening that sharply increase b-ion fragment coverage, raising sequence-identification confidence and improving de novo sequencing. Also supports 9-plex stable-isotope multiplexing at ~240 samples/day, boosting protein data points roughly 6.5x over the unlabelled baseline in sensitive (low-input) proteomics.

Applied in

General proteomics, Venomics

Papers describing them (49)

Authors (208)

Adelina Rogowska‐Wrzesinska, Aeran Jeon, Ahmet Yesilcimen, Albert T. Lebedev, Almary Chacon, Amela Hozić, Ana Butorac, Andrea Janeš, Andrew Ellington, Anick Girard, Ariane Borges, Arundhati Nag, Arye Tishbee, Atsushi Kitanaka, Ayaka Hayashi, Bart Devreese, Bart Samyn, Bella DiChiara, Bhanuramanand Kuchibhotla, Bongjin Moon, Bradley L. Pentelute, Caroline Wang, Charles W. Johannes, Cheng Guo, Chi Nguyen, Chihiro Nakajima, Christien Kluwe, Corinna Friedrich, Cunlong Zhang, Daisuke Nakayama, Dandan Chen, Daniel C. Liebler, David Barnes-Seeman, Dhaval Nanavati, Dongmei Zhang, Douglas A. Gage, Douglas S. Masterson, Dragan Gamberger, Dukjin Kang, Eiji Ando, Ema Svetličić, Emily Atkinson, Eric Biron, Fumiya Honda, Gianluca Picariello, Giuseppe Raucci, Griet Debyser, Han Bin Oh, Harrison Specht, Henna Patel, Henry Shion, Heramb M. Kulkarni, Hie-Joon Kim, Hiroaki Suga, Hiroki Kuyama, Hiroyuki Awane, Hisashi Miyagawa, Hongxia Liu, Hui Luan, Huiyong Yin, Hye‐Ki Min, Hyuksu Kwon, Inae Jang, Ingemar Lindh, Iryna Onasenko, Ivana Dodig, J. Gregory Zeikus, Jagannadham V. Medicherla, James A. Madsen, Jan Sjövall, Janko Diminić, Jasna Mrvčić, Jeffrey J. Wilson, Jennifer S. Brodbelt, Jia Sun, Jin Wang, Jin Yong Neo, Jin-Su Song, Jingyu Moon, Joe R. Cannon, John Allison, John C. Gebler, Jong-Seo Kim, Jozef Van Beeumen, Jue Chen, Jumpei Morimoto, Jungjoo Nam, Kaishun Bi, Karlo Hock, Kasa Shiva Raju, Kazuhiro Sonomura, Keisuke Shima, Kevin McDonnell, Khiry L Patterson, Kjell Sergeant, Konstantin A. Artemenko, Lars Hjelmqvist, Lei Zhao, Li Quan Koh, Lucija Dončević, Luka Ozdanovac, M.V. Jagannadham, Maddy Yeh, Madhusudhana Rao Nalam, Madison Letendre, Mansup Shin, Mao Yakio, Marija Nišavić, Mario Cindrić, Maris Laivenieks, Mark Adamo, Martin P. Lacey, Masahiro Miyashita, Meliha Solak Mekić, Meri Gabrielli, Michael Alan Lee, Michael P. Agius, Michelle R. Robinson, Minoru Yamaguchi, Min‐Sik Kim, Mitsuru Tanaka, Ned A. Porter, Ngoc K. Nguyen, Nicholas Ellor, Nikita B. Poljakov, Nikolai Slavov, Noa Petler, Norikazu Ueyama, Osamu Nishimura, Paolo Conrotto, Peter J. Lee, Peter Roepstorff, Ping Chen, R. S. Youngquist, R. Srinivas, Rachel Shu Ting Chan, Renata Biba, Rene Kandler, Risa Katagihara, Ryotaro Asai, Sabrina Novelli, Samir Das, Samy Memmi, Sang Yun Han, Sang‐Won Lee, Sankara Rao Kola, Sarah Sipe, Saya Nakamura, Scott A. Robotham, Sergey V. Kovalev, Seung Bum Park, Shangfu Li, Shigemi Norioka, Shinsuke Sando, Shusheng Zhang, Song Nie, Songhie An, Song‐Ping Liang, Stephen H. Collins, Sun Young Lee, Susumu Tsunasawa, Swamy V. Cherukuvada, T. Keough, Takashi Nakazawa, Takashi Obama, Taka‐aki Okamura, Tamara Allakhverdova, Tatiana Yu. Samgina, Tomas Bergman, Toru Yoshikawa, Toshiro Matsui, Tsz Ying Yuen, Tun-Li Shen, Ulf Hellman, V. Ramesh, Vishnu M. Dhople, Višnja Bačun‐Družina, Vladimir Gorshkov, Wai Kit Pang, Wayne Vuong, Wei Mi, Weibin Chen, William J. Griffiths, Woo‐Suk Choie, Xianchun Wang, Xiangming Hu, Xiaohui Chen, Xiaolan Chen, Xinxia Liang, Xuefeng Guo, Xumin Zhang, Yanchang Huang, Yee Hwee Lim, Yi Wee Lim, Yizhi Xiao, Yoichiro Nihashi, Yong Ho Lee, Yongju Kim, Yoshiaki Nakagawa, Yosuke Hanai, Yuki Goto, Yumiko Toyama, Yutaro Saito, Yuyang Jiang, Yuzo Yamazaki, Zachary P. Gates, Zhi-Heng Huang, Zhi’ang Chen

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