PMID- 17105210
OWN - NLM
STAT- MEDLINE
DCOM- 20070124
LR  - 20190816
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 45
IP  - 46
DP  - 2006 Nov 21
TI  - Dissection of a human septin: definition and characterization of distinct domains 
      within human SEPT4.
PG  - 13918-31
AB  - The septins are a conserved family of guanosine-5'-triphosphate (GTP)-binding 
      proteins. In mammals they are involved in a variety of cellular processes, such 
      as cytokinesis, exocytosis, and vesicle trafficking. Specifically, SEPT4 has also 
      been shown to be expressed in both human colorectal cancer and malignant 
      melanoma, as well as being involved in neurodegenerative disorders. However, many 
      of the details of the modes of action of septins in general remain unclear, and 
      little is known of their detailed molecular architecture. Here, we define 
      explicitly and characterize the domains of human SEPT4. Regions corresponding to 
      the N-terminal, GTPase, and C-terminal domains as well as the latter two together 
      were successfully expressed in Escherichia coli in soluble form and purified by 
      affinity and size-exclusion chromatographies. The purified domains were analyzed 
      by circular dichroism spectroscopy, fluorescence spectroscopy, dynamic light 
      scattering, and small-angle X-ray scattering, as well as with bioinformatics 
      tools. Of the three major domains that comprise SEPT4, the N-terminal domain 
      contains little regular secondary structure and may be intrinsically 
      unstructured. The central GTPase domain is a mixed alpha/beta structure, probably 
      based on an open beta sheet. As defined here, it is catalytically active and 
      forms stable homodimers in vitro. The C-terminal domain also forms homodimers and 
      can be divided into two regions, the second of which is alpha-helical and 
      consistent with a coiled-coil structure. These studies should provide a useful 
      basis for future biophysical studies of SEPT4, including the structural basis for 
      their involvement in diseases such as cancer and neurodegenerative disorders.
FAU - Garcia, Wanius
AU  - Garcia W
AD  - Centro de Biotecnologia Molecular e Estrutural (CBME), Instituto de Fisica de Sao 
      Carlos (IFSC), Universidade de Sao Paulo (USP), Av. Trabalhador Sao Carlense 400, 
      centro, Box 369, Sao Carlos, SP, 13560-970, Brazil. wanius@if.sc.usp.br
FAU - de Araujo, Ana Paula Ulian
AU  - de Araujo AP
FAU - Neto, Mario de Oliveira
AU  - Neto Mde O
FAU - Ballestero, Michel R M
AU  - Ballestero MR
FAU - Polikarpov, Igor
AU  - Polikarpov I
FAU - Tanaka, Manami
AU  - Tanaka M
FAU - Tanaka, Tomoo
AU  - Tanaka T
FAU - Garratt, Richard Charles
AU  - Garratt RC
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (Cytoskeletal Proteins)
RN  - EC 3.6.1.- (GTP Phosphohydrolases)
RN  - EC 3.6.1.- (SEPTIN4 protein, human)
RN  - EC 3.6.1.- (Septins)
SB  - IM
MH  - Amino Acid Sequence
MH  - Chromatography, Affinity
MH  - Chromatography, Gel
MH  - Circular Dichroism
MH  - Cytoskeletal Proteins/chemistry/genetics/*metabolism
MH  - Electrophoresis, Polyacrylamide Gel
MH  - GTP Phosphohydrolases/chemistry/genetics/*metabolism
MH  - Humans
MH  - Molecular Sequence Data
MH  - Protein Structure, Secondary
MH  - Septins
MH  - Sequence Homology, Amino Acid
MH  - Spectrometry, Fluorescence
EDAT- 2006/11/16 09:00
MHDA- 2007/01/25 09:00
CRDT- 2006/11/16 09:00
PHST- 2006/11/16 09:00 [pubmed]
PHST- 2007/01/25 09:00 [medline]
PHST- 2006/11/16 09:00 [entrez]
AID - 10.1021/bi061549z [doi]
PST - ppublish
SO  - Biochemistry. 2006 Nov 21;45(46):13918-31. doi: 10.1021/bi061549z.