• 主页
  • 查找结果
  • The large intracellular loop of hZIP4 is an intrinsically disordered zinc binding domain.

The large intracellular loop of hZIP4 is an intrinsically disordered zinc binding domain.

Metallomics : integrated biometal science (2015-04-18)
Elizabeth M Bafaro, Sagar Antala, Tuong-Vi Nguyen, Stephen P Dzul, Brian Doyon, Timothy L Stemmler, Robert E Dempski
摘要

The human (h) ZIP4 transporter is a plasma membrane protein which functions to increase the cytosolic concentration of zinc. hZIP4 transports zinc into intestinal cells and therefore has a central role in the absorption of dietary zinc. hZIP4 has eight transmembrane domains and encodes a large intracellular loop between transmembrane domains III and IV, M3M4. Previously, it has been postulated that this domain regulates hZIP4 levels in the plasma membrane in a zinc-dependent manner. The objective of this research was to examine the zinc binding properties of the large intracellular loop of hZIP4. Therefore, we have recombinantly expressed and purified M3M4 and showed that this domain binds two zinc ions. Using a combination of site-directed mutagenesis, metal binding affinity assays, and X-ray absorption spectroscopy, we demonstrated that the two Zn(2+) ions bind sequentially, with the first Zn(2+) binding to a CysHis3 site with a nanomolar binding affinity, and the second Zn(2+) binding to a His4 site with a weaker affinity. Circular dichroism spectroscopy revealed that the M3M4 domain is intrinsically disordered, with only a small structural change induced upon Zn(2+) coordination. Our data supports a model in which the intracellular M3M4 domain senses high cytosolic Zn(2+) concentrations and regulates the plasma membrane levels of the hZIP4 transporter in response to Zn(2+) binding.

材料
货号
品牌
产品描述

Sigma-Aldrich
甘油, for molecular biology, ≥99.0%
Sigma-Aldrich
羟乙基哌嗪乙硫磺酸, ≥99.5% (titration)
Sigma-Aldrich
羟乙基哌嗪乙硫磺酸, BioPerformance Certified, ≥99.5% (titration), suitable for cell culture
Sigma-Aldrich
甘油, ≥99.5%
Sigma-Aldrich
IPTG, ≥99% (TLC), ≤0.1% Dioxane
Sigma-Aldrich
3-吗啉丙磺酸, ≥99.5% (titration)
Sigma-Aldrich
乙二胺四乙酸, ACS reagent, 99.4-100.6%, powder
Sigma-Aldrich
焦碳酸二乙酯, ≥97% (NMR)
Sigma-Aldrich
乙二胺四乙酸, anhydrous, BioUltra, ≥99% (titration)
Sigma-Aldrich
乙二醇-双(2-氨基乙醚)-N,N,N′,N′-四乙酸, for molecular biology, ≥97.0%
Sigma-Aldrich
HEPES缓冲溶液, 1 M in H2O
Sigma-Aldrich
甘油, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for electrophoresis, ≥99% (GC)
Sigma-Aldrich
N -乙基马来酰亚胺, crystalline, ≥98% (HPLC)
Sigma-Aldrich
乙二醇-双(2-氨基乙醚)-N,N,N′,N′-四乙酸, ≥97.0%
Sigma-Aldrich
乙二胺四乙酸, anhydrous, crystalline, BioReagent, suitable for cell culture
Sigma-Aldrich
乙二胺四乙酸 溶液, 0.02% in DPBS (0.5 mM), sterile-filtered, BioReagent, suitable for cell culture
SAFC
羟乙基哌嗪乙硫磺酸
Sigma-Aldrich
3-吗啉丙磺酸, BioPerformance Certified, suitable for cell culture, ≥99.5% (titration)
Sigma-Aldrich
乙二胺四乙酸, purified grade, ≥98.5%, powder
Sigma-Aldrich
甘油, meets USP testing specifications
Sigma-Aldrich
甘油, BioXtra, ≥99% (GC)
Sigma-Aldrich
甘油 溶液, 83.5-89.5% (T)
Sigma-Aldrich
焦碳酸二乙酯, 99% (NT)
Sigma-Aldrich
异丙 β-D-1-硫代半乳糖苷, ≥99% (TLC)
Sigma-Aldrich
羟乙基哌嗪乙硫磺酸, BioUltra, for molecular biology, ≥99.5% (T)
Sigma-Aldrich
N -乙基马来酰亚胺, BioUltra, ≥99.0% (HPLC)
Sigma-Aldrich
乙二胺四乙酸, 99.995% trace metals basis
Sigma-Aldrich
甘油, FCC, FG
Sigma-Aldrich
甘油, BioUltra, for molecular biology, anhydrous, ≥99.5% (GC)
Sigma-Aldrich
异丙基 β-D-硫代半乳糖吡喃糖苷 溶液, ReadyMade IPTG solution for Blue-white screening