Enhanced sequence coverage, better identification of combinatorial co-occurring PTMs and improved detection of proteoforms are key highlights of middle-down approach and hence, this can be a promiscuous approach for protein sequencing and proteomics.

Sabareesh V

Centre for Bio-Separation and Technology

Vellore Campus

Pandeswari P.B

Fulltext

Vellore Institute of Technology (VIT) is a private university located in&nbsp;Tamil Nadu, India. Founded in 1984, as Vellore Engineering College, the institution offers 20 undergraduate, 34 postgraduate, four integrated and four research programs. It has campuses in Vellore, Amravati, Bhopal and Chennai.

VIT is one of the top ranked private universities in India according to NIRF, THE and QS Rankings.&nbsp;Govt. of India has recognized&nbsp;VIT, Vellore as an&nbsp;Institution of Eminence. This has allowed VIT to take independent quality initiatives and move up in world ranking.

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VIT University

Middle-down approach: a choice to sequence and characterize proteins/proteomes by mass spectrometry

RSC Advances

The potential of immobilized metal/chelate affinity (IMA) in a continuous fashion, referred as conjoint approach, to pre-fractionate plasma proteins (in their native state) prior to LC–MS analysis was investigated in this study. Four transition metal-ions (Co (II), Zn (II), Ni (II) and Cu (II)) were individually chelated with IDA (iminodiacetic acid) coated CIM (Convective Interaction Media) disks and placed in a single housing in the following sequential order: IDA-Co (II) → IDA-Zn (II) → IDA-Ni (II) → IDA-Cu (II). The rationale behind this order is to retain proteins based on their specific requirement for surface exposed histidine topography. This structural pre-fractionation hypothesis was successfully proven using four human plasma proteins (fibrinogen, IgG, transferrin, and albumin) with varying histidine topographies. This conjoint IMA pre-fractionation strategy not only fractionated proteins (from plasma) based on their native surface histidine topography, but also identified 157 proteins from human plasma. The advantage of our conjoint IMA is its ability to fractionate proteins in their native state and reduce plasma complexity in a single step by employing single buffer system. © 2017 Elsevier B.V.

Journal of Chromatography B

A structure based plasma protein pre-fractionation using conjoint immobilized metal/chelate affinity (IMA) system

Affinity selection of histidine-containing peptides using metal chelate methacrylate monolithic disk for targeted LC–MS/MS approach in high-throughput proteomics

Mass Spectrometry Reviews

Anatomy and evolution of database search engines—a central component of mass spectrometry based proteomic workflows

The Skyline ecosystem: Informatics for quantitative mass spectrometry proteomics

Bioinformatics

VisioProt-MS: interactive 2D maps from intact protein mass spectrometry

Journal	Data powered by TypesetRSC Advances
Publisher	Data powered by TypesetRoyal Society of Chemistry (RSC)
Open Access	No