title: Determination of membrane protein molecular weights and association equilibrium constants using sedimentation equilibrium and sedimentation velocity.
abstract: Regulated molecular interactions are essential for cellular function and viability, and both homo- and hetero-interactions between all types of biomolecules play important cellular roles. This chapter focuses on interactions between membrane proteins. Knowing both the stoichiometries and stabilities of these interactions in hydrophobic environments is a prerequisite for understanding how this class of proteins regulates cellular activities in membranes. Using examples from the authors' work, this chapter highlights the application of analytical ultracentrifugation methods in the determination of these parameters for integral membrane proteins. Both theoretical and practical aspects of carrying out these experiments are discussed.
authors.affiliation: T. C. Jenkins Department of Biophysics, Johns Hopkins University, Baltimore, Maryland 21218, USA.
authors.forename: Nancy K
authors.initials: NK
authors.lastname: Burgess
authors.forename: Ann Marie
authors.initials: AM
authors.lastname: Stanley
authors.forename: Karen G
authors.initials: KG
authors.lastname: Fleming
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000302:isolation & purification / Q000378:metabolism*; D019281:Dimerization; D004926:Escherichia coli / Q000378:metabolism*; D007700:Kinetics; D008970:Molecular Weight; D054466:Phospholipases A1 / Q000302:isolation & purification / Q000378:metabolism*; D018272:Porins / Q000302:isolation & purification / Q000378:metabolism*; D013816:Thermodynamics; D014461:Ultracentrifugation / Q000379:methods*
chemical_list: D001425:Bacterial Outer Membrane Proteins; C109237:OmpF protein; D018272:Porins; D054466:Phospholipases A1; C412876:outer membrane phospholipase A