Soybean Proteomics - Plant Proteomics: Methods and Protocols - page 306

Table 1
(continued)

Conditions Cultivar Organ/organelle Proteomic methodologies Reference

Ozone Enrei Leaf (chloroplast) IEF tube gel, SDS-PAGE,

MALDI-TOF MS, protein
sequencing

[ 24 ]

Infection by Williams 82 Root hairs IPG, gradient acrylamide gel,

MALDI-TOF MS, Q-TOF
MS

[ 37 ]

B. japonicum Eb-b0-1

En1282
Enrei

Root IPG, SDS-PAGE,

nanoLC-MS/MS

[ 16 ]

Seed fi lling Maverick Developing seed IPG, SDS-PAGE, MALDI-

TOF MS

[ 23 ]

Maverick Developing seed IPG, SDS-PAGE, nESI-

LC-MS/MS

[ 26 ]

Storage
protein

Jefferson Seed IPG, SDS-PAGE, MALDI-

TOF MS

[ 22 ]

Developmental
stages

Enrei Leaf/fl ower IEF tube gel, SDS-PAGE,

MALDI-TOF MS, protein
sequencing

[ 12 ]

pathways in mesophyll cells indicate the presence of secondary
phenolic metabolism in soybean leaves [ 8 ]. Presence of these inter-
fering substances not only hampers high-quality protein extraction
[ 9 ], but also impedes protein spot separation in high resolution
2-DE gels, resulting in streaking, smearing, and a signifi cant reduc-
tion in the number of distinctly resolved protein spots [ 10 ]. Instead
of these limitations, much progress has been made in standardiza-
tion of the protein extraction methodologies from different tissues
of soybean. Natarajan et al. [ 11 ] compared four different protein
extraction/solubilization methods—urea, thiourea/urea, phenol,
and a modifi ed TCA /acetone to determine their effectiveness in
separating soybean seed proteins by 2-DE. The thiourea/urea and
TCA methods were found to be more suitable in resolving less
abundant and high molecular weight proteins. In addition, these
two methods exhibited higher protein resolution and spot inten-
sity as compared to rest of the methods.
To compare proteomic changes of soybean leaves and fl owers
at various developmental stages, Ahsan and Komatsu [ 12 ] evalu-
ated three different protein extraction protocols—TCA precipita-
tion [ 13 ], phenol extraction method [ 14 ] with modifi cations and
direct tissue homogenizing in suitable protein solubilization buf-
fers. To optimize protein pellet solubilization buffer, A-buffer [ 1 ]