Analysis of Peptides & Proteins / PPSQ

Applications for PPSQ

 
 

Protein Analysis Platform: Combining the powerful capabilities of MALDI-TOF MS (MALDI-8020) and Edman Sequencing (PPSQ-50A Gradient System) for accurate N-terminal sequence of peptides

Mass spectrometry has become an indispensable tool for researchers looking to sequence peptides. Although effective in many cases, sequencing by In Source Decay (ISD) faces a few challenges its ability to provide reliable sequence information including isobaric amino acids, database dependency and low molecular weight interferences. Traditional Edman sequencing avoids mass dependency and the use of databases by analyzing each amino acid from the N-terminus one at a time in sequence. Unfortunately, Edman has its own limitations in providing high sequence coverage. This technical note investigates the benefits of combining the intact mass and sequencing information from the MALDI-8020 (ISD) with the N-terminal sequence obtained from the PPSQ-50A gradient system (Edman). Using the combined information allows investigators the ability to obtain a more complete picture of their proteins and peptides of interest.

 
 
 
 
 
 
 
 
 

Primary Structural Analysis of Proteins / Peptides – Application of Protein Sequencer –

Understanding of pathological states and life phenomena has accelerated dramatically with recent widespread advances in the field of proteomics, which encompasses the exhaustive comparison and identification of expressed proteins in normal and disease-state tissues and cells. The scope of this investigation includes pathology, differences in type, genetic variation and cell abnormalities across a range of scientific disciplines. For example, investigation of the causes of disease and treatment methods, and the search for specific biomarkers associated with those diseases in the field of proteomics has grown rapidly. The word "proteomics" was coined in the early 1990s to indicate the entire set of proteins expressed in a specific cell type or organism associated with that genome. In general, there is very little in the way of change in the genome in a given type of organism, but the change in the proteome between organs and cells can be vast. In order for an organism to make its genetic information actually function, the process of gene expression is required. 

 
 
 
 
 
 
 
 
 

N-terminal Amino Acid Sequencing of Mouse IgG Using the PPSQ-51A/53A Gradient System

As genome analysis technology has evolved, the genome database has become more comprehensive, and proteome analysis which is one form of post-genome analysis using the database - has grown by leaps and bounds. As a result of technical developments in mass spectrometers, we were able to achieve high-throughput analysis in the amino acid sequencing of proteins by using a mass spectrometer and its database. Although many proteins in the genome database are registered as precursor proteins, the various proteins expressed in cells are transported into the body as mature proteins that result from processing of these precursor proteins.

 
 
 
 
 
 
 
 
 

Amino Acid Sequence Analysis of Peptides and Proteins with Modified Amino Acid Using PPSQ Isocratic System

Protein identification with a mass spectrometer (MS) and search engine utilizing genomic databases has now become the main stream in analysis of proteins. Although the proteins in the genomic databases are registered as precursor proteins, the expressed proteins in living cells are modified after translation and have various functions. However, since there are differences in the theoretical mass number of the precursor protein and the mature protein modified after translation, the score and reliability of search results obtained by the MS analysis and search engine is sometimes low. Moreover, identification of amino acid sequences by MS without using databases is both quite complex and difficult. On the other hand, a protein sequencer using the conventional Edman degradation method obtains highly reliable sequencing results, and amino acid sequences can be identified easily even in case the database is inadequate. 

 
 
 
 
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