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Protein cell biology studies how proteins function within biological cells: where they are delivered, which partners they encounter, and how these interactions affect cellular processes. A protein's protein primary structure and abundance do not by themselves specify its location or available partners. RNA-binding proteins, G proteins and protein targeting illustrate different aspects of that problem.

The microsomal protein import experiment makes location experimentally testable. In 1975, Günter Blobel and Bernhard Dobberstein combined cell-free translation components with dog-pancreas microsomes. An immunoglobulin light chain product made while suitable membranes were present became processed and resistant to external proteolysis, whereas globin did not enter the same protected compartment. Adding membranes only after synthesis did not reproduce the effect in this preparation.^[ref]

The important observation is controlled accessibility, not just recovery near a membrane. A protein can bind a vesicle's outside without crossing it; enclosure can protect it from an external protease. Protease protection assays therefore complement measurements of total product and membrane topology.

For an original two-compartment example, suppose a fixed total of100 competent cargo molecules is divided between20 enclosed and80 exposed molecules. Redistributing the same total to70 enclosed and30 exposed changes accessibility even though total abundance stays100. If every enclosed molecule is protected and every exposed molecule is digested in the stipulated model, the protected fraction changes from0.20 to0.70. Real measurements also need checks for membrane integrity, protease activity and product recovery; the fractions alone do not identify every transport mechanism. This is why cellular organization belongs alongside the isolated molecule's chemistry.

Protein targeting directs a protein to its cellular destination. Synthesis in the cytosol does not imply that the protein works there.

For entry into the endoplasmic reticulum, distinguish three questions: does the nascent chain reach the membrane, does part of it cross into the ER lumen, and does a segment remain embedded as a transmembrane protein? Cotranslational protein translocation couples these events to biological translation. A signal peptide can initiate delivery without specifying the eventual destination of the mature protein.

This distinction matters experimentally. Finding a protein in a membrane-containing centrifugation pellet could reflect surface binding or protein aggregation. The microsomal protein import experiment instead asks whether an intact membrane prevents an external protease from reaching the product. That tests accessibility across a barrier, rather than proximity to it.

A signal peptide is a short targeting segment of a newly made polypeptide chain. A familiar ER signal sequence lies near the N-terminus and is removed during entry; Günter Blobel and Bernhard Dobberstein's 1975 experiments linked a larger immunoglobulin light chain precursor to a processed product.^[ref]

Cleavage is a separate question from recognition. Not every targeting signal is a cleaved signal peptide: an internal signal anchor can remain part of a transmembrane protein. Conversely, loss of a segment does not by itself prove membrane crossing. Compare the independent size and accessibility readouts in the microsomal protein import experiment.