Ion Mobility-Mass Spectrometry Reveals Highly-Compact Intermediates in the Collision Induced Dissociation of Charge-Reduced Protein Complexes.
Protocols that aim to construct complete models of multiprotein complexes based on ion mobility and mass spectrometry data are becoming an important element of integrative structural biology efforts. However, the usefulness of such data is predicated, in part, on an ability to measure individual subunits removed from the complex while maintaining a compact/folded state. Gas-phase dissociation of intact complexes using collision induced dissociation is a potentially promising pathway for acquiring such protein monomer size information, but most product ions produced are possessed of high charge states and elongated/string-like conformations that are not useful in protein complex modeling. It has previously been demonstrated that the collision induced dissociation of charge-reduced protein complexes can produce compact subunit product ions; however, their formation mechanism is not well understood. Here, we present new experimental evidence for the avidin (64 kDa) and aldolase (157 kDa) tetramers that demonstrates significant complex remodeling during the dissociation of charge-reduced assemblies. Detailed analysis and modeling indicates that highly compact intermediates are accessed during the dissociation process by both complexes. Here, we present putative pathways that describe the formation of such ions, as well as discuss the broader significance of such data for structural biology applications moving forward.
Department of Chemistry, University of Michigan, Ann Arbor, MI, 48109, USA.
Russell E
RE
Bornschein
Department of Chemistry, University of Michigan, Ann Arbor, MI, 48109, USA.
Shuai
S
Niu
Department of Chemistry, University of Michigan, Ann Arbor, MI, 48109, USA.
Joseph
J
Eschweiler
Department of Chemistry, University of Michigan, Ann Arbor, MI, 48109, USA. bruotolo@umich.edu.
Brandon T
BT
Ruotolo
D000818:Animals; D001360:Avidin; D002645:Chickens; D005634:Fructose-Bisphosphate Aldolase; D007477:Ions / Q000032:analysis / Q000737:chemistry*; D008958:Models, Molecular; D046912:Multiprotein Complexes / Q000032:analysis / Q000737:chemistry*; D011817:Rabbits; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D007477:Ions; D046912:Multiprotein Complexes; D001360:Avidin; D005634:Fructose-Bisphosphate Aldolase
Collision induced unfolding; Electrospray ionization; Ion-ion chemistry; Noncovalent complex; Structural biology