PDBsum entry 2kvg

Transcription

PDB id: 2kvg

Contents

Protein chain

27 a.a.

Metals

_ZN

PDB id:

2kvg

Name:

Transcription

Title:

Structure of the three-cys2his2 domain of mouse testis zinc finger protein

Structure:

Zinc finger and btb domain-containing protein 32. Chain: a. Fragment: mtzd_finger3, c2h2-type 3 domain, unp residues 402-428. Synonym: repressor of gata, testis zinc finger protein. Engineered: yes

Source:

Mus musculus. Mouse. Organism_taxid: 10090. Expressed in: escherichia coli. Expression_system_taxid: 562.

NMR struc:

15 models

Authors:

C.-C.Chou,Y.-C.Lou,C.Chen

Key ref:

C.C.Chou et al. (2010). Structure and DNA binding characteristics of the three-Cys(2)His(2) domain of mouse testis zinc finger protein. Proteins, 78, 2202-2212. PubMed id: 20544958 DOI: 10.1002/prot.22732

Date:

15-Mar-10 Release date: 14-Apr-10

Protein chain

Q9JKD9  (ZBT32_MOUSE) -  Zinc finger and BTB domain-containing protein 32 from Mus musculus

Seq: 465 a.a.

Struc: 27 a.a.

Enzyme reactions

• Enzyme class: E.C.?

DOI no: 10.1002/prot.22732

Proteins 78:2202-2212 (2010)

PubMed id: 20544958

Structure and DNA binding characteristics of the three-Cys(2)His(2) domain of mouse testis zinc finger protein.

C.C.Chou, Y.C.Lou, T.K.Tang, C.Chen.

ABSTRACT

The C-terminal three-Cys(2)His(2) zinc-finger domain (TZD) of mouse testis zinc-finger protein binds to the 5'-TGTACAGTGT-3' at the Aie1 (aurora-C) promoter with high specificity. Interestingly, the primary sequence of TZD is unique, possessing two distinct linkers, TGEKP and GAAP, and distinct residues at presumed DNA binding sites at each finger, especially finger 3. A K(d) value of approximately 10(-8) M was obtained from surface plasmon resonance analysis for the TZD-DNA complex. NMR structure of the free TZD showed that each zinc finger forms a typical beta beta alpha fold. On binding to DNA, chemical shift perturbations and the R(2) transverse relaxation rate in finger 3 are significantly smaller than those in fingers 1 and 2, which indicates that the DNA binding affinity in finger 3 is weaker. Furthermore, the shift perturbations between TZD in complex with the cognate DNA and its serial mutants revealed that both ADE7 and CYT8, underlined in 5'-ATATGTACAGTGTTAT-3', are critical in specific binding, and the DNA binding in finger 3 is sequence independent. Remarkably, the shift perturbations in finger 3 on the linker mutation of TZD (GAAP mutated to TGEKP) were barely detected, which further indicates that finger 3 does not play a critical role in DNA sequence-specific recognition. The complex model showed that residues important for DNA binding are mainly located on positions -1, 2, 3, and 6 of alpha-helices in fingers 1 and 2. The DNA sequence and nonsequence-specific bindings occurring simultaneously in TZD provide valuable information for better understanding of protein-DNA recognition.