The fusion of intracellular transport vesicles with their target membranes requires the assembly of SNARE proteins anchored in the apposed membranes. Here we use recombinant cytoplasmic domains of the yeast SNAREs involved in Golgi to plasma membrane trafficking to examine this assembly process in vitro. Binary complexes form between the target membrane SNAREs Sso1p and Sec9p; these binary complexes can subsequently bind to the vesicle SNARE Snc2p to form ternary complexes. Binary and ternary complex assembly are accompanied by large increases in alpha-helical structure, indicating that folding and complex formation are linked. Surprisingly, we find that binary complex formation is extremely slow, with a second-order rate constant of approximately 3 M(-1) s(-1). An N-terminal regulatory domain of Sso1p accounts for slow assembly, since in its absence complexes assemble 2,000-fold more rapidly. Once binary complexes form, ternary complex formation is rapid and is not affected by the presence of the regulatory domain. Our results imply that proteins that accelerate SNARE assembly in vivo act by relieving inhibition by this regulatory domain.

Despite the unique, sleep-disrupting occupational demands of military personnel, in-person and Internet CBTi are efficacious treatments for this population. The effect sizes for in-person were consistently better than Internet and both were similar to those found in civilians. Dissemination of CBTi should be considered for maximum individual and population benefits, possibly in a stepped-care model.

Evaluation of new respiratory symptoms in military personnel after service in Southwest Asia should focus on airway hyperreactivity from exposures to higher levels of ambient particulate matter. These patients may be difficult to diagnose and require close follow-up.

Registry: ClinicalTrials.gov; Title: Comparing Internet and In-Person Brief Cognitive Behavioral Therapy of Insomnia; Identifier: NCT01549899; URL: https://clinicaltrials.gov/ct2/show/NCT01549899.

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