pressin acts on the inner medullary collecting duct (IMCD) in the kidney to regulate water and urea transport. To obtain a "parts list" of gene products expressed in the IMCD, we carried out mRNA profiling of freshly isolated rat IMCD cells using Affymetrix Rat 230 2.0 microarrays with ϳ31,000 features; 7,913 annotated transcripts were found to be expressed above background in the IMCD cells. We have created a new online database (the "IMCD Transcriptome Database;" http://dir.nhlbi.nih.gov/papers/lkem/imcdtr/) to make the results publicly accessible. Among the 30 transcripts with the greatest signals on the arrays were 3 water channels: aquaporin-2, aquaporin-3, and aquaporin-4, all of which have been reported to be targets for regulation by vasopressin. In addition, the transcript with the greatest signal among members of the solute carrier family of genes was the UT-A urea transporter (Slc14a2), which is also regulated by vasopressin. The V2 vasopressin receptor was strongly expressed, but the V1a and V1b vasopressin receptors did not produce signals above background. Among the 200 protein kinases expressed, the serumglucocorticoid-regulated kinase (Sgk1) had the greatest signal intensity in the IMCD. WNK1 and WNK4 were also expressed in the IMCD with a relatively high signal intensity, as was protein kinase A (␤-catalytic subunit). In addition, a large number of transcripts corresponding to A kinase anchoring proteins and 14-3-3 proteins (phospho-S/T-binding proteins) were expressed. Altogether, the results combine with proteomics studies of the IMCD to provide a framework for modeling complex interaction networks responsible for vasopressin action in collecting duct cells. oligonucleotide array; vasopressin; aquaporin-2; urea transport; kinases THE INNER MEDULLARY COLLECTING DUCT (IMCD) is the terminal portion of the collecting duct system of the kidney. The collecting duct system represents the final site of adjustment of urinary composition and volume and therefore is critical for extracellular fluid homeostasis. An important regulator of collecting duct transport function is vasopressin, which controls both water and urea transport (21,53).To address the mechanisms of vasopressin signaling in the renal IMCD, we have been following a "systems-biology" approach consisting of identification of the component proteins via protein mass spectrometry-based analysis (2,29,30,62,71,102) and antibody-based quantification of protein abundance (41) coupled to computational analysis of the identified proteins to discover signaling networks involved in IMCD regulation (30).

Correct phosphorylation site assignment is a critical aspect of phosphoproteomic analysis. Large-scale phosphopeptide data sets that are generated through liquid chromatography-coupled tandem mass spectrometry (LC-MS/MS) analysis often contain hundreds or thousands of phosphorylation sites that require validation. To this end, we have created PhosphoScore, an open-source assignment program that is compatible with phosphopeptide data from multiple MS levels (MS(n)). The algorithm takes into account both the match quality and normalized intensity of observed spectral peaks compared to a theoretical spectrum. PhosphoScore produced >95% correct MS(2) assignments from known synthetic data, > 98% agreement with an established MS(2) assignment algorithm (Ascore), and >92% agreement with visual inspection of MS(3) and MS(4) spectra.

Querying uncertain data sets (represented as probability distributions) presents many challenges due to the large amount of data involved and the difficulties comparing uncertainty between distributions. The Earth Mover's Distance (EMD) has increasingly been employed to compare uncertain data due to its ability to effectively capture the differences between two distributions. Computing the EMD entails finding a solution to the transportation problem, which is computationally intensive. In this paper, we propose a new lower bound to the EMD and an index structure to significantly improve the performance of EMD based Knearest neighbor (K-NN) queries on uncertain databases.We propose a new lower bound to the EMD that approximates the EMD on a projection vector. Each distribution is projected onto a vector and approximated by a normal distribution, as well as an accompanying error term. We then represent each normal as a point in a Hough transformed space. We then use the concept of stochastic dominance to implement an efficient index structure in the transformed space. We show that our method significantly decreases K-NN query time on uncertain databases. The index structure also scales well with database cardinality. It is well suited for heterogeneous data sets, helping to keep EMD based queries tractable as uncertain data sets become larger and more complex.

Opsins are a large group of proteins with seven transmembrane segments (TMSs) that are found in all domains of life. There are two types of opsins that are sometimes considered nonhomologous: type I is known from prokaryotes and some eukaryotes, while type II is known only from Eumetazoan animals. Type II opsins are members of the family of G-protein coupled receptors (GPCRs), which facilitate signal transduction across cell membranes. While previous studies have concluded that multiple transmembrane-containing protein families-including type I opsins-originated by internal domain duplication, the origin of type II opsins has been speculated on but never tested. Here we show that type II opsins do not appear to have originated through a similar internal domain duplication event. This provides further evidence that the two types of opsins are nonhomologous, indicating a convergent evolutionary origin, in which both groups of opsins evolved a seven-TM structure and light sensitivity independently. This convergence may indicate an important role for seven-TM protein structure for retinal-based light sensitivity.