Recent Research Articles from UNTHSC

Syndicate content NCBI pubmed
NCBI: db=pubmed; Term="University of North Texas Health Science Center"[All Fields] OR "Univ. of North Texas Health Science Center"[All Fields] OR "UNT Health Science Center"[All Fields] OR "Osteopathic Research Center"[All Fields] OR "University of North Texas System College of Pharmacy"[All Fields] OR "UNT System College of Pharmacy"[All Fields] OR "College of Pharmacy, University of North Texas System"[All Fields]
Updated: 2 hours 30 min ago

Detergent screening of the human voltage-gated ion channel (Hv1) using fluorescence-detection size exclusion chromatography.

Sat, 05/31/2014 - 4:07am
Related Articles

Detergent screening of the human voltage-gated ion channel (Hv1) using fluorescence-detection size exclusion chromatography.

Protein Sci. 2014 May 23;

Authors: Agharkar A, Rzadkowolski J, McBroom M, Gonzales EB

Abstract
The human voltage-gated proton channel (Hv1) is a membrane protein consisting of four transmembrane domains and internal amino- and carboxy-termini. The protein is activated by membrane depolarization, similar to other voltage-sensitive proteins. However, the Hv1 proton channel lacks a traditional ion pore. The human Hv1 proton channel has been implicated in mediating sperm capacitance, stroke, and most recently as a biomarker/mediator of cancer metastasis. Recently, the three-dimensional structures for homologues of this voltage-gated proton channel were reported. However, it is not clear what artificial environment is needed to facilitate the isolation and purification of the human Hv1 proton channel for structural study. In the present study, we generated a chimeric protein that placed an enhanced green fluorescent protein (EGFP) to the amino-terminus of the human Hv1 proton channel (termed EGFP-Hv1). The chimeric protein was expressed in a baculovirus expression system using Sf9 cells and subjected to detergent screening using fluorescence-detection size exclusion chromatography (FSEC). The EGFP-Hv1 proton channel can be solubilized in the zwitterionic detergent Anzergent 3-12 and the nonionic n-dodecyl-β-D-maltoside (DDM) with little protein aggregation and a prominent monomeric protein peak at 48 hours post infection. Furthermore, we demonstrate that the chimeric protein exhibits a monomeric protein peak, which is distinguishable from protein aggregates, at the final size exclusion chromatography purification step. Taken together, we can conclude that solubilization in DDM will provide a useable final product for further structural characterization of the full-length human Hv1 proton channel.

PMID: 24863684 [PubMed - as supplied by publisher]

Editorial comment: Symposium: 2012 Musculoskeletal Infection Society.

Sat, 05/31/2014 - 4:07am
Related Articles

Editorial comment: Symposium: 2012 Musculoskeletal Infection Society.

Clin Orthop Relat Res. 2013 Oct;471(10):3098-9

Authors: Nana A, Wongworawat MD

PMID: 23836244 [PubMed - indexed for MEDLINE]

Genome-guided discovery of diverse natural products from Burkholderia sp.

Sat, 05/31/2014 - 4:07am
Related Articles

Genome-guided discovery of diverse natural products from Burkholderia sp.

J Ind Microbiol Biotechnol. 2014 Feb;41(2):275-84

Authors: Liu X, Cheng YQ

Abstract
Burkholderia species have emerged as a new source of diverse natural products. This mini-review covers all of the natural products discovered in recent years from Burkholderia sp. by genome-guided approaches--these refer to the use of bacterial genome sequence as an entry point for in silico structural prediction, wet lab experimental design, and execution. While reliable structural prediction based on cryptic biosynthetic gene cluster sequence was not always possible due to noncanonical domains and/or module organization of a deduced biosynthetic pathway, a molecular genetic method was often employed to detect or alter the expression level of the gene cluster to achieve an observable phenotype, which facilitated downstream natural product purification and identification. Those examples of natural product discovery from Burkholderia sp. provide practical guidance for future exploration of Gram-negative bacteria as a new source of natural products.

PMID: 24212473 [PubMed - indexed for MEDLINE]