Protein–protein Interactions: Identification

Abstract

Protein–protein interactions are essential features of biological processes that allow cells to grow, divide and respond to their environment. A variety of methodologies now exist to probe protein–protein interactions in vivo and in vitro.

Keywords: protein–protein interactions; oligomerization; affinity tag; co‐immunoprecipitation; protein complex

Figure 1.

(a) A. tumefaciens Tram homodimer; (b) vesicular stomatitis virus G protein homotrimer; (c) chimaeric heterotrimeric G protein; (d) obligatory homodimerization of α globin and β globin in human haemoglobin; (e) nonobligatory heterodimer of human growth hormone complexed with human growth hormone receptor.

Figure 2.

Affinity tag pulldown experiments allow selective capture of a tagged protein (red circles) and binding partners (green circles) on to a solid support, such as a bead. Co‐immunoprecipitation experiments are based on similar principles, except that antibodies are used to tether bait proteins to a solid support.

Figure 3.

(a) Fluorophores (green circles) excited with polarized light emit polarized photons (black arrows emanating from fluorophores). Measuring fluorescence emission with polarizations parallel and perpendicular to the polarization of excitation provides an indication of the degree to which the fluorescently labelled protein molecules (blue circles) are tumbling in solution; (b) protein‐binding partners (orange circles) interact with the fluorescently labelled proteins to form a complex that tumbles more slowly, increasing the polarization and the anisotropy of the fluorophores.

Figure 4.

(a) To achieve fluorescence resonance energy transfer, the donor emission spectrum must overlap with the acceptor absorption spectrum; (b) a protein with a donor fluorophore (cyan circle) is excited with photons of wavelength λ1 and emits photons of wavelength λ2. When brought into close proximity to an acceptor fluorophore (yellow circle) due to protein complex formation, FRET results in emission by the acceptor fluorophore of photons of wavelength λ3.

close

References

Causier B (2004) Studying the interactome with the yeast two‐hybrid system and mass spectrometry. Mass Spectrometry Reviews 23(5): 350–367.

Chen G, Malenkos JW, Cha MR, Fuqua C and Chen L (2004) Quorum‐sensing antiactivator TraM forms a dimer that dissociates to inhibit TraR. Molecular Microbiology 52(6): 1641–1651.

Clackson T, Ultsch MH, Wells JA and de Vos AM (1998) Structural and functional analysis of the 1:1 growth hormone:receptor complex reveals the molecular basis for receptor affinity. Journal of Molecular Biology 277(5): 1111–1128.

Fields S and Song O (1989) A novel genetic system to detect protein–protein interactions. Nature 340(6230): 245–246.

Frier JA and Perutz MF (1977) Structure of human foetal deoxyhaemoglobin. Journal of Molecular Biology 112(1): 97–112.

Lambright DG, Sondek J, Bohm A et al. (1996) The 2.0 A crystal structure of a heterotrimeric G protein. Nature 379(6563): 311–319.

Nooren IM and Thornton JM (2003) Diversity of protein–protein interactions. EMBO Journal 22(14): 3486–3492.

Puig O, Caspary F, Rigaut G et al. (2001) The tandem affinity purification (TAP) method: a general procedure of protein complex purification. Methods 24(3): 218–229.

Roche S, Bressanelli S, Rey FA and Gaudin Y (2006) Crystal structure of the low‐pH form of the vesicular stomatitis virus glycoprotein G. Science 313(5784): 187–191.

Further Reading

Golemis E (ed.) (2002) Protein–Protein Interactions : A Molecular Cloning Manual. New York: Cold Spring Harbor Laboratory Press.

Mitchell JC (2005) Identification of Protein–Protein Interactions. Encyclopedia of Life Sciences. Chichester: Wiley.

Contact Editor close
Submit a note to the editor about this article by filling in the form below.

* Required Field

How to Cite close
Lopper, Matthew E, and Keck, James L(Jul 2007) Protein–protein Interactions: Identification. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0020491]