| MadSci Network: Cell Biology |
1. Actually, the cell really is extremely crowded. Your library will
likely have a copy of "The Machinery of Life" by David S. Goodsell
(Springer-Verlag, 1992). Goodsell draws cells and cell components to
scale and based on real data. The cover alone ought to convince you how
crowded a cell is.
Now to protein trafficking.
2. Soluble proteins. Soluble proteins destined for the cytosol are
simply synthesized and possibly directly released off the endoplasmic
reticulum (ER). Those that are modified by addition of carbohydrate or
lipid may sometime go to the Golgi apparatus through the soluble interior
of the ER which is directly connected to the soluble space of the Golgi.
However, they can usually be modified directly by soluble enzymes.
3. Membrane proteins. Membrane proteins are inserted into the ER
membrane AS THEY ARE BEING MADE. They achieve their final
conformation/topology while in the ER, or at least a topology very close
to final. They then move laterally through the ER membrane to the Golgi.
This likely involves interactions with some set of cytoskeletal components
that act as attachment points and motors. Once in the Golgi, they get
modified if that's what that particular protein needs.
How do the various membrane proteins get to their respective
organelles? For the most part, membrane proteins appear to have specific
short amino acids sequences that direct that protein to a particular
organ. For example, a sequence for translocation to the nucleus is 5 a.a.
in length and contains all aspartates and glutamates. Other signals are
present which will send a protein to a mitochondrion or back to the ER,
etc.
In the Golgi, proteins bound, for example, to the mitochondrion end
up being sorted out as they pass through the various layers of the Golgi
until they are together. Once together and when they have reached the
final membranes of the Golgi, they "bud" off as a vesicle. The
localization signal of the proteins appears to somehow tell the machinery
to take that vesicle to the mitocondrion where it fuses and thus inserts
new protein into the membrane. Localization signals may or may not
(usually) be cleaved off.
Switching the short localization sequence on a protein from, for
example, nucleus to mitochondrial will cause that protein to be directed
to the wrong organ where it may or may not function properly or possibly
even be toxic.
4. Now finally to bacteria. Membrane proteins in a bacterium are
inserted into the membrane DURING SYNTHESIS. If, for example, amino acids
10-30 form a membrane domain, that segment is being inserted into the
membrane by the time the ribosome has attached a.a. 40 or 45. For insight
into the insertion process, see Heinrich et al., Cell (2000) 102:233-244.
How are they sorted? Good question. We don't know for most proteins
but the answer I think everyone assumes is that localization to a
particular part of the cell is due to specific protein-protein
interactions. This is however quite dynamic and localization can change
very quickly.
For insertion into the outer membrane, the same machinery that inserts
proteins into the plasma/cell membrane probably takes outer membrane
proteins also but there is an additional component(s) (probably) that help
guide insertion into the outer membrane (of the Gram negatives). I
suspect that the machinery used for this purpose has somehow been coopted
by evolution to form the Type III secretion systems (there are 4 systems
known) that pathogenic bacteria use to insert proteins not only through
their inner and outer membranes but directly into the plasma membrane or
even into the cytosol of the cell they are invading. For a general review
of secretion systems use PubMed or some other search program and find
reviews by Tony Pugsley. For some insight into the various specific
proteins and some good pictures, see papers by Joe Lutkenhaus, Piet de
Boer, Janine Maddock and Lucy Shapiro in addition to Pugsley.
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