Article (Scientific journals)
Multi-Enzymatic Limited Digestion: The Next-Generation Sequencing for Proteomics?
Morsa, Denis; Baiwir, Dominique; La Rocca, Raphaël et al.
2019In Journal of Proteome Research
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Keywords :
PTMs; de novo sequencing; digestibility profile; limited digestion; mAbs; mass spectrometry; protein characterization; proteolytic method
Abstract :
[en] Over the past 40 years, proteomics, generically defined as the field dedicated to the identification and analysis of proteins, has tremendously gained in popularity and potency through advancements in genome sequencing, separative techniques, mass spectrometry, and bioinformatics algorithms. As a consequence, its scope of application has gradually enlarged and diversified to meet specialized topical biomedical subjects. Although the tryptic bottom-up approach is widely regarded as the gold standard for rapid screening of complex samples, its application for precise and confident mapping of protein modifications is often hindered due to partial sequence coverage, poor redundancy in indicative peptides, and lack of method flexibility. We here show how the synergic and time-limited action of a properly diluted mix of multiple enzymes can be exploited in a versatile yet straightforward protocol to alleviate present-day drawbacks. Merging bottom-up and middle-down ideologies, our results highlight broad assemblies of overlapping peptides that enable refined and reliable characterizations of proteins, including variant identification, and their carried modifications, including post-translational modifications, truncations, and cleavages. Beyond this boost in performance, our methodology also offers efficient de novo sequencing capabilities, in view of which we here present a dedicated custom assembly algorithm.
Disciplines :
Chemistry
Author, co-author :
Morsa, Denis  ;  Université de Liège - ULiège > Département de chimie (sciences) > Chimie analytique inorganique
Baiwir, Dominique  ;  Université de Liège - ULiège > GIGA Platforms
La Rocca, Raphaël  ;  Université de Liège - ULiège > Département de chimie (sciences) > Laboratoire de spectrométrie de masse (L.S.M.)
Zimmerman, Tyler A.
Hanozin, Emeline ;  Université de Liège - ULiège > Département de chimie (sciences) > Laboratoire de spectrométrie de masse (L.S.M.)
Grifnée, Elodie ;  Université de Liège - ULiège > Département de chimie (sciences) > Laboratoire de spectrométrie de masse (L.S.M.)
Longuespée, Rémi ;  Université de Liège - ULiège > Département de chimie (sciences) > Chimie analytique inorganique
Meuwis, Marie-Alice  ;  Centre Hospitalier Universitaire de Liège - CHU > Service de gastroentérologie, hépatologie, onco. digestive
Smargiasso, Nicolas 
De Pauw, Edwin  ;  Université de Liège - ULiège > Département de chimie (sciences) > Chimie analytique inorganique
Mazzucchelli, Gabriel  ;  Université de Liège - ULiège > Département de chimie (sciences) > Laboratoire de spectrométrie de masse (L.S.M.)
Language :
English
Title :
Multi-Enzymatic Limited Digestion: The Next-Generation Sequencing for Proteomics?
Publication date :
2019
Journal title :
Journal of Proteome Research
ISSN :
1535-3893
eISSN :
1535-3907
Publisher :
American Chemical Society, Washington, United States - District of Columbia
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 24 May 2019

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