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Legume Genomics and Genetics (online), 2011, Vol. 2, No.1, 1-5
http://lgg.sophiapublisher.com
1
Research Article Open Access
Isotypes of 78 kD streptavidin biotin-binding proteins (SBP) during initial ger-
mination of dark-grown pea (
Pisum sativum
L.) seedlings
Mahmoud F.M. Moustafa
1,2
1. Botany Department, Faculty of Science, South Valley University, Egypt
2. Department of Biological Sciences, Faculty of Science, King Khalid University, Abha, Kingdom of Saudi Arabia
Corresponding author email: hamdony@yahoo.com;
Authors
Legume Genomics and Genetics 2011, Vol.2 No.1 doi:10.5376/lgg.2011.02.0001
Received: 20 Feb., 2011
Accepted: 12 Mar., 2011
Published: 16 Mar., 2011
This is an Open Access article published under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and
reproduction in any medium, provided the original work is properly cited.
Preferred citation for this article as:
Moustafa M.F.M., 2011, Isotypes of 78 kD streptavidin biotin-binding proteins (SBP) during initial germination of dark-grown pea (
Pisum sativum
L.)
seedlings, Legume Genomics and Genetics (online), Vol.2, No.1 pp.1-5 (doi: 10.5376/lgg.2011.02.0001)
Abstract
In this study two dimensional page and western blotting have been used to investigate the status of isotypes of 78 kD
streptavidin biotin-binding proteins (SBP) in dry seeds, germinated seeds and a combination with its different plant parts including
stems, plumules and roots of pea (
Pisum sativum
L. var Alaska). Dry seeds contain 21 isotypes, five of them are almost separated
with isoelectric point (pI) 5.66, 5.82, 5.88, 5.91 and 6.55, and four of them are partially separated with p
I
values 5.92, 5.94, 5.96 and
5.98, the remaining forming unseparated isotypes mixture with pI values varied from 5.07 to 6.50. Embryos of forty hours germinated
pea seeds showed 9 different biotin isotypes with pI values from 5.76 to 6.64. Stems parts of seventy two hours germinated pea seeds
showed highest expressed 11 biotin isotypes with pI values from 5.60 to 6.65, whereas the plumules parts showed 10 isotypes with pI
values from 5.78 to 6.46. And the root parts showed 5 separated isotypes with P
I
values from 5.72 to 6.00 and unseparated isotypes
mixture with a pI values ranged from 6.27 to 6.67. These results suggest that the isotypes of 78 kD streptavidin biotin-binding
proteins in dormant and germinating pea seeds might play a role in embryo longevity and early seed germination.
Keywords
Isoelectric point; Isotypes;
Pisum sativum
L.
Background
Streptavidin and avidin are one of the most successful
and widely used as detection reagents in immunology,
biochemistry and cell biology due to their high affinity
binding to biotin. Biotin found in all organisms is an
essential cofactor for a small number of enzymes
necessary for CO
2
transfer during carboxylation reactions
(Knowles, 1989). In plant cells biotin-dependent
enzyme is required for both growth of vegetative tissues
and synthesis of storage lipids in developing pea seed
(Stumpf, 1980; Harwood, 1988). In yeast (
Pichia
pastoris
) biotin deficiency results in lower growth
rates and lower biomass yields in batch cultures and in
wash-out in continuous cultures (Jungo, 2007). In
animals cells biotin-containing enzymes catalyse
ATP-dependent CO
2
fixation and are thus important
enzymes in intermediary metabolism (Dakshinamurti
and Chauhan, 1989). Furthermore, stored biotin in
muscle is of special interest because alterations in
mitochondrial biotin-containing enzymes are likely to
be involved in most metabolic myopathies (Scholte et
al., 1987). In bacteria such as
Escherichia coli
,
Bacillus
subtilis
, and
Bacillus sphaericus
, biotin biosynthesis
has been elucidated by combined biochemical and
genetic studies (Marquet et al., 2001). In higher plants
initial genetic information on biotin synthesis came
from analysis of the
bio1
biotin auxotroph of Arabidopsis
that lead to identification of the second biotin auxotroph
(Schneider et al., 1989).
It is reported that in pea seeds only one type of seed-
specific biotinylated-protein and three biotin-dependent
plant carboxylases (Wurtele and Nikolau, 1990; Duval
et al., 1993; Alban et al., 1993; Harwood 1988). And
animals cells contain four biotinylated enzyme acetyl-
CoA carboxylase (ACC; EC 6.4.1.2), 3-methy-lcrotonyl-
CoA carboxylase (MCC; EC 6.4.1.4), pro- pionyl-CoA
carboxylase (PCC; EC 6.4.1.3) and pyruvate car-
boxylase (PC; EC 6.4.1.1) (Zempleni and Mock,