MECHANISM OF

PROTEIN TARGETING
INTO ENDOPLASMIC
RETICULUM
REBATI RAMAN PANDA
PART-II
BOTANY DEPARTMENT,
BERHAMPUR UNIVERSITY
 INTRODUCTION
Protein targeting or protein sorting is the mechanism by which a cell
transports proteins to the appropriate positions in the cell or outside of it.
•DNA synthesis maintains the genetic information and passes this to the
next generation
•RNA synthesis (transcription) is a transfer of the information from the
DNA where it is stored into RNA which can be transported and
interpreted.
•Ribosomes translate the nucleotides on the mRNA into amino acid
sequences producing a polypeptide
TRANSLATION
● Initiation – the assembly of a ribosome on an
mRNA molecule.
● Elongation – repeated cycles of amino acid
addition.
● Termination – the release of the new protein
chain.
 PROTEIN TARGETING
Both in prokaryotes and eukaryotes, newly synthesized proteins must be delivered to a specific
subcellular location or exported from the cell for correct activity. This phenomenon is called
protein targeting.
• Protein targeting is necessary for proteins that are destined to work outside the cytoplasm.
• This delivery process is carried out based on information contained in the protein itself.
• Correct sorting is crucial for the cell; errors can lead to diseases .
PROTEIN TRANSLOCATION
• In 1970, Günter Blobel conducted experiments on the translocation of proteins across
membranes.
• He was awarded the 1999 Nobel Prize for his findings. He discovered that many proteins
have a signal sequence, that is, a short amino acid sequence at one end that functions like a
postal code for the target organelle.
MAJOR PROTEIN - SORTING
PATHWAYS

1. Secretory pathway

2. Non-secretory
pathway
Targeting pathways
 MAJOR
QUSTIONS
1.What is the nature of the targeting sequence, and what
distinguishes it from other types of targeting sequences?
2.What is the receptor for the targeting sequence?
3.What is the structure of the translocation channel that
allows transfer of proteins across the membrane bilayer?
In particular, is the channel so narrow that proteins can
pass through only in an unfolded state, or will it
accommodate folded protein domains?
4.What is the source of energy that drives unidirectional
transfer across the membrane?
Synthesis of secreted
proteins, integral plasma-
membrane proteins, and
proteins destined for the
ER, Golgi complex, or
lysosome begins on
cytosolic ribosomes,
which become attached to
the membrane of the ER,
forming the rough ER
THE SIGNAL SEQUENSES OF
DIFFERENT SECRECATORY PROTINS
ALL CONTAIN ONE OR MORE +VE
CHARGED AMINOACID ADJACENT
TO A CONTINOUS STRETCH OF 6-12
HAYDROPHOBIC RECIDUE ALSO
KNOWN AS HYDROPHOBIC CORE.
THIS HYDROPHOBIC CORE DIRECTS
THE NASCENT SECRETORY PROTINE
TO THE ER.
 STRUCTURE OF A SIGNAL
RECOGNITION PARTICLE (SRP)
Sec61α is a translocon component. Cross-linking experiments
show that Sec61α is a translocon component that contacts
nascent secretory proteins as they pass into the ER lumen.
An mRNA encoding the N-terminal 70 amino acids of the secreted
protein prolactin was translated in a cell-free system containing
microsomes (see Figure 13-4b). The mRNA lacked a chain-
termination codon and contained one lysine codon, near the
middle of the sequence. The reaction mixtures contained a
chemically modified lysyl-tRNA in which a light-activated cross-
linking reagent was attached to the lysine side chain. Although the
entire mRNA was translated, the completed polypeptide could not
be released from the ribosome without a chain- termination
codon and thus became “stuck” crossing the ER membrane. The
reaction mixtures were then exposed to intense light, which
caused the nascent polypeptide chain to become covalently
bound to whatever proteins were near it in the translocon. When
the experiment was performed using microsomes from
mammalian cells, the nascent chain became covalently linked to
Sec61α. Different versions of the prolactin mRNA were created so
that the modified lysine residue would be placed at different
distances from the ribosome; cross-linking to Sec61α was
observed only when the modified lysine was positioned within the
translocation channel.
 TOPOLOGICAL CLASSES
OF INTIGRAL MEMBREN
• PROTINon the rough ER include five topological
Proteins synthesized
classes of integral membrane proteins as well as a lipid-
anchored type.
• Topogenic sequences—N-terminal signal sequences, internal
stop-transfer anchor sequences, and internal signal- anchor
sequences—direct the insertion of nascent proteins into the
ER membrane and their orientation within it. This orientation
is retained during transport of the completed membrane
protein to its final destination—e.g., the plasma membrane
 REFERENCES
• Lodish H., Berk A., Kaiser C.A., Krieger M., Bretscher
A., Ploegh H., Amon A., Martin K.C., Molecular cell
biology (8th edition), Page: 583-600

• Cooper, G.M., The cell; A molecular approach (7th

• Protein translocation across the endoplasmic reticulum