Protein oligomerization monitored by fluorescence fluctuation spectroscopy: self-assembly of rubisco activase.

ABSTRACT: A methodology is presented to characterize complex protein assembly pathways by fluorescence correlation spectroscopy. We have derived the total autocorrelation function describing the behavior of mixtures of labeled and unlabeled protein under equilibrium conditions. Our modeling approach allows us to quantitatively consider the relevance of any proposed intermediate form, and K(d) values can be estimated even when several oligomeric species coexist. We have tested this method on the AAA+ ATPase Rubisco activase (Rca). Rca self-association regulates the CO(2) fixing activity of the enzyme Rubisco, directly affecting biomass accumulation in higher plants. However, the elucidation of its assembly pathway has remained challenging, precluding a detailed mechanistic investigation. Here, we pre

SUBMITTER: Chakraborty M 

PROVIDER: S-EPMC3433602 | biostudies-literature | 2012 Sep

REPOSITORIES: biostudies-literature