Functional interrogation of the kinome using nucleotide acyl phosphates.

The central role of protein kinases in signal transduction pathways has generated intense interest in targeting these enzymes for a wide range of therapeutic indications. Here we report a method for identifying and quantifying protein kinases in any biological sample or tissue from any species. The procedure relies on acyl phosphate-containing nucleotides, prepared from a biotin derivative and ATP or ADP. The acyl phosphate probes react selectively and covalently at the ATP binding sites of at least 75% of the known human protein kinases. Biotinylated peptide fragments from labeled proteomes are captured and then sequenced and identified using a mass spectrometry-based analysis platform to determine the kinases present and their relative levels. Further, direct competition between the probes and inhibitors can be assessed to determine inhibitor potency and selectivity against native protein kinases, as well as hundreds of other ATPases. The ability to broadly profile kinase activities in native proteomes offers an exciting prospect for both target discovery and inhibitor selectivity profiling.

Relationship between depression and psychosocial functioning after traumatic brain injury.

OBJECTIVE: To examine the relationship between depression and psychosocial functioning up to 5 years after traumatic brain injury (TBI). DESIGN: Longitudinal cohort study with 2 assessments completed. SETTING: Community. PARTICIPANTS: Individuals (N=188) with TBI living in the community. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Structured Clinical Interview for Depression, self-reports of depression severity, functional symptoms, quality of life (QOL), unmet important needs, and psychosocial functioning. RESULTS: Based on observed depression patterns at initial and repeat assessment, 4 subgroups were created: no depression, resolved depression, late-onset depression, and chronic depression. Groups were equivalent in terms of demographic and injury-related factors but differed significantly in perceived psychosocial functioning. The no-depression group reported fewer depressive symptoms and higher levels of psychosocial functioning, whereas the chronic-depression group reported the poorest psychosocial functioning, with a further decline in QOL at reassessment. Although the resolved-depression and late-onset-depression groups reported similar psychosocial functioning at initial assessment, psychosocial functioning had improved for the resolved-depression group and declined for the late-onset-depression group at reassessment. Pre- and postpsychiatric diagnoses were common in all groups, with pre-TBI diagnosis of depression not predictive of post-TBI depression. CONCLUSIONS: Findings highlight the need for broad-based assessments and timely interventions for both mood and psychosocial challenges after TBI.