SoyProLow: A protein database enriched in low abundant soybean proteins.

Mona Tavakolan,Savithiry S Natarajan,Benjamin F Matthews,Nadim W Alkharouf

doi:10.6026/97320630010599

Abstract

Soybeans are an important legume crop that contain 2 major storage proteins, β-conglycinin and glycinin, which account about 70- 80% of total seed proteins. These abundant proteins hinder the isolation and characterization of several low abundant proteins in soybean seeds. Several protein extraction methodologies were developed in our laboratory to decrease these abundant storage proteins in seed extracts and to also decrease the amount of ribulose-1, 5-bisphosphate carboxylase/oxygenase (RuBisCO), which is normally very abundant in leaf extracts. One of the extraction methodologies used 40% isopropanol and was more effective in depleting soybean storage proteins and enhancing low abundant seed proteins than similar methods using 10-80% isopropanol. Extractions performed with 40% isopropanol decreased the amount of storage proteins and revealed 107 low abundant proteins when using the combined approaches of two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and Mass Spectrometry (MS). The separation of proteins was achieved by iso-electric focusing (IEF) and 2D-PAGE. The proteins were analyzed with MS techniques to provide amino acid sequence. The proteins were identified by comparing their amino acid sequences with those in different databases including NCBI-non redundant, UniprotKB and MSDB databases. In this investigation, previously published results on low abundant soybean seed proteins were used to create an online database (SoyProLow) to provide a data repository that can be used as a reference to identify and characterize low abundance proteins. This database is freely accessible to individuals using similar techniques and can be for the subsequent genetic manipulation to produce value added soybean traits. An intuitive user interface based on dynamic HTML enables users to browse the network and the profiles of the low abundant proteins.Availabilityhttp://bioinformatics.towson.edu/Soybean_low_abundance_proteins_2D_Gel_DB/Gel1.aspx

Highlights

Soybean (Glycine max) seeds provide an inexpensive source of amylase, cytochrome c, and urease
These two compounds account for 80 % of total introduction of silencing genes, or to responses to various proteins and are responsible for much of the nutritional, physicochemical and physiological properties of soybean seeds
Protein extraction for iso-electric focusing (IEF) is difficult because seed samples have lipids and other compounds that disrupt or interfere with focusing during protein separation steps

Summary

Introduction

Soybean (Glycine max) seeds provide an inexpensive source of amylase, cytochrome c, and urease. Soybean seeds contain numerous proteins that provide nutrients to the embryo and protect the seed from diseases and pests. Soybean seeds contain two major storage proteins, β -conglycinin and Bowman-Birk Inhibitors (BBI) of chymotrypsin and trypsin [1, 2].

Results

Conclusion