Biosynthesis of storage Proteins

Jeeva.V
2023533004
M.Sc,. Horticulture
 Biosynthesis of Storage Proteins
• Storage Proteins function in storing amino acids as nutrients and as

building blocks for the growing organisms.

• It can also be found in root and shoot tubers like in potatoes.

• The major storage proteins in pulses is globulins and prolamins in

cereals.
• Plant storage proteins were systematically studied and categorized by

T.B. Osborne on the basis of their solubility characters as

– Albumins – water soluble

– Globulins – soluble in dilute salt solution

– Prolamins – soluble in 70-80% alcohol

– Glutelins – soluble in dilute acids or alkalies

Albumins
 Albumins are readily soluble in water, dilute acids and alkalies.
 Coagulated by heat.
 Seed proteins contain albumin in lesser quantities.
 Albumins may be precipitated out from solution using high salt
concentration, a process called 'salting out'.
 They are deficient in glycine.
 Serum albumin and ovalbumin (egg white) are examples.
Globulins
 Globulins are insoluble or sparingly soluble in water, but their solubility
is greatly increased by the addition of neutral salts such as sodium
chloride.
These proteins are coagulated by heat.
 They are deficient in methionine.
 Serum globulin, fibrinogen, myosin of muscle and globulins of pulses are
examples.
 Types of globulins:

α1 globulin: e.g. antitrypsin

α2 globulin: e.g. hepatoglobin: protein that binds hemoglobin to
prevent its excretion by the kidney
β-globulin: e.g. transferrin: protein that transport iron
γ-globulins: Immunoglobulins (antibodies): responsible for immunity.
• Globulins are the most widespread group of storage protein
present mainly in dicot seeds
• Globulins are divided into two groups based on their
sedimentation coefficient – 7S Vicilins and 11S Legumins.
• 7S vicilins are glycosylated trimeric proteins and lack cysteine
residues and do not form disulfide bonds.
• Mature 11S legumins consists of six subunits pairs, each pair
consisting of an acidic and a basic polypeptide.
• 11S legumins are very rarely glycosylated.
Prolamins
 Prolamins are insoluble in water but soluble in 70-80% aqueous alcohol.
 Upon hydrolysis they yield much proline and amide nitrogen, hence the
name prolamin.
 They are deficient in lysine.
 Gliadin of wheat and zein of corn are examples of prolamins.
 Prolamins are unique that they occur only in endosperms of cereals and
other grasses.
 Eg. are rice and oats (Oryzenin of rice, Gliadin of wheat, Hordein of barley,
Avenin of oats, Zein of maize)
 N-terminal domain – proline and glutamine
 In cereals they act as trypsin inhibitors, amylase or both
 Legume albumins functions as enzyme inhibitors (hetero dimer linked by
disulphide bonds
Glutelins
 Glutelins are insoluble in water and absolute

alcohol but soluble in dilute alkalies and acids.

 They are plant proteins e.g., glutenin of wheat.
 Biosynthesis of storage proteins
• Storage proteins are coded by specific genes located in the nuclear DNA.
• Transcription of the genes to mRNA occurs in the nucleus of the cell.
• mRNA is synthesized only in the specific tissues and within a discrete
period during seed development.
• Prolamin genes are transcribed only in the starchy endosperm of cereals
but not in the aleurone layer which is also part of the endosperm.
• The mRNA produced by transcription undergoes splicing and forms
mature processed mRNA.
• The mature mRNA is transported from the nucleus to the rough
endoplasmic reticulum (RER) where the protein synthesis (translation)
occurs in the ribosomes.
• The primary translation product is called preproprotein.
• A preproprotein contain a signal sequence at the N-terminal end which
directs the transport of the newly translated polypeptides into the lumen of
RER.
• In lumen of RER signal sequences is removed and other post-translational
modifications occur.
• Finally, the protein is folded into three dimensional structure.
 Synthesis of 11S legumin
• Steps in the synthesis of 11S legumin (glycinin) in soybean cotyledons is
shown in the figure.
• The transcription product of legumin gene is an unprocessed mRNA
containing nontranslated introns, which are spliced out to produce the
mature, processed legumin mRNA.
• The mRNA is transported from the nucleus to the rough endoplasmic
reticulum (RER) where it is translated.
• The primary translation product (preprolegumin) contains an acidic (A)
and a basic (B) subunit joined by a 4-amino acid linker sequence.
• It also contains a signal sequence at the N-terminal and a pentapeptide at
the C-terminal end.
• The signal peptide and the pentapeptide are cleaved and the polypeptide
assembles into trimers.
• The trimers enter the protein bodies.
• Mature hexameric legumin assembles after removal of the linker peptide.
• Protein body formation occurs in two different ways in developing cereal grain

endosperms.

• They can be formed either from vacuole or from endoplasmic reticulum.

• The protein synthesized in the endoplasmic reticulum will be transported to the

vacuole by way of golgi apparatus and the protein bodies form by fragmentation of

the vacuole.

• 11S globulins in endosperm of rice and oats; globulins and albumins in aleurone

layer of cereals are formed by this way.

• Prolamins of rice and maize are sequestered in the lumen of the endoplasmic

reticulum and form localized distensions of protein bodies.

• The rice endosperm thus contains two populations of protein bodies, those of

vacuolar origin, which contain globulin-like protein, glutelin (80% of storage

protein) and those of ER origin, which contains prolamin (oryzenin) (5% of storage

protein).
Biosynthesis of storage proteins
 Post Translational Modification of Proteins

Amino-Terminal and Carboxyl-Terminal Modifications

Loss of Signal Sequences
Modification of Individual Amino Acids
Attachment of Carbohydrate Side Chains
Addition of Isoprenyl Groups
Addition of Prosthetic Groups
Proteolytic Processing
Formation of Disulfide Cross-Links
Role of Signal sequence
 Hypothetical
protein folding
pathway
Much of secondary
structure is present
No tertiary structure
-- Molten globule

Hydrophobic
collapsed state is
known as molten
globule
Linear pathway for
folding a two-domain
protein
A simulated protein folding pathway

Short stretches of
secondary
Unfolded
structure
Has many possible
conformations

Folded
Native conformation
 Levinthal’s paradox
Consider a protein of 100 amino acids. Assign 2 conformations to each amino acid.
The total conformations of the protein is 2100=1.27x1030. Allow 10-13 sec for the
protein to sample through one conformation in search for the overall energy
minimum. The time it needs to sample through all conformations is

(10-13)(1.27x1030)=1.27x1017sec = 4x109 years!

Levinthal’s paradox illustrates that proteins must only sample through limited
conformations, or fold by “specific pathways”. Much research efforts are devoted
in searching for the principles of the “specific pathways.”.
 Free-energy funnel description of
the protein folding pathway
 Folding Accesory Proteins

A. Protein disulfide isomerases PDI

B. Peptidyl Proline isomerases - Catalyses cis-trans isomerisation of

peptide bonds involving proline
C. Molecular chaperones - Help folding, especially of large proteins, by
preventing interaction with other proteins
Protein folding by Chaperons
Protein folding by Chaperons
Forces that stabilizes the Protein