N-terminal amino acid sequence determination of proteins by N-terminal dimethyl labeling: Pitfalls and advantages when compared with Edman degradation sequence analysis Journal Article

   

Authors:	Chang, E.; Pourmal, S.; Zhou, C.; Kumar, R.; Teplova, M.; Pavletich, N. P.; Marians, K. J.; Erdjument-Bromage, H.
Article Title:	N-terminal amino acid sequence determination of proteins by N-terminal dimethyl labeling: Pitfalls and advantages when compared with Edman degradation sequence analysis
Abstract:	In recent history, alternative approaches to Edman sequencing have been investigated, and to this end, the Association of Biomolecular Resource Facilities (ABRF) Protein Sequencing Research Group (PSRG) initiated studies in 2014 and 2015, looking into bottom-up and top-down N-terminal (Nt) dimethyl derivatization of standard quantities of intact proteins with the aim to determine Nt sequence information.We have expanded this initiative and used low picomole amounts of myoglobin to determine the efficiency of Nt-dimethylation. Application of this approach on protein domains, generated by limited proteolysis of overexpressed proteins, confirms that it is a universal labeling technique and is very sensitive when compared with Edman sequencing. Finally, we compared Edman sequencing and Nt-dimethylation of the same polypeptide fragments; results confirm that there is agreement in the identity of the Nt amino acid sequence between these 2 methods. © 2016 ABRF.
Keywords:	bottom-up ms; in-gel digestion; nt protein dimethylation; nt sequencing
Journal Title:	Journal of Biomolecular Techniques
Volume:	27
Issue:	2
ISSN:	1524-0215
Publisher:	ABRF
Date Published:	2016-07-01
Start Page:	61
End Page:	74
Language:	English
DOI:	10.7171/jbt.16-2702-002
PROVIDER:	scopus
PMCID:	PMC4802820
PUBMED:	27006647
DOI/URL:	https://www.scopus.com/inward/record.uri?eid=2-s2.0-84977497374&partnerID=40&md5=993b3f7b6be8bc0c6ccb6b6a6b982cc9
Notes:	Article -- Export Date: 2 August 2016 -- Source: Scopus

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