Recombinant expression, purification and characterisation of the HMG domain of human SRY

被引：12
作者：
Kelly, S
Yotis, J
Macris, M
Harley, V
机构：
[1] Prince Henrys Inst Med Res, Clayton, Vic 3168, Australia
[2] Univ Melbourne, Dept Biochem, Melbourne, Vic 3010, Australia
[3] Univ Melbourne, Howard Florey Inst Expt Physiol & Med, Melbourne, Vic 3010, Australia
来源：
PROTEIN AND PEPTIDE LETTERS | 2003年 / 10卷 / 03期
关键词：
HMG; SRY HMG domain; SOX; recombinant protein expression; FPLC purification; mass spectrometry; DNA binding; calmodulin binding;
D O I：
10.2174/0929866033479004
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
The HMG domain is a DNA binding and bending 'architectural' motif involved in chromatin re-modelling during transcription. Recombinant SRY HMG domain protein, 88 amino acids in length, has been produced in E coli. Using FPLC and a stirred ultra-filtration cell, this domain has been purified to homogeneity and concentrated to yield milligram quantities. Functional characterisation studies of the pure, concentrated SRY HMG domain show the recombinantly expressed protein to be active in terms of DNA binding and calmodulin binding activities.
引用
收藏
页码：281 / 286
页数：6
相关论文
共 8 条
[1]   Phylogeny of the SOX family of developmental transcription factors based on sequence and structural indicators [J].
Bowles, J ;
Schepers, G ;
Koopman, P .
DEVELOPMENTAL BIOLOGY, 2000, 227 (02) :239-255
[2]   SRY, LIKE HMG1, RECOGNIZES SHARP ANGLES IN DNA [J].
FERRARI, S ;
HARLEY, VR ;
PONTIGGIA, A ;
GOODFELLOW, PN ;
LOVELLBADGE, R ;
BIANCHI, ME .
EMBO JOURNAL, 1992, 11 (12) :4497-4506
[3]   Heparin elution of transcription factors from DNA-Sepharose columns [J].
Gadgil, H ;
Jarrett, HW .
JOURNAL OF CHROMATOGRAPHY A, 1999, 848 (1-2) :131-138
[4]   DNA-BINDING ACTIVITY OF RECOMBINANT SRY FROM NORMAL MALES AND XY FEMALES [J].
HARLEY, VR ;
JACKSON, DI ;
HEXTALL, PJ ;
HAWKINS, JR ;
BERKOVITZ, GD ;
SOCKANATHAN, S ;
LOVELLBADGE, R ;
GOODFELLOW, PN .
SCIENCE, 1992, 255 (5043) :453-456
[5]   The HMG box of SRY is a calmodulin binding domain [J].
Harley, VR ;
LovellBadge, R ;
Goodfellow, PN ;
Hextall, PJ .
FEBS LETTERS, 1996, 391 (1-2) :24-28
[6]   The DNA-binding specificity of SOX9 and other SOX proteins [J].
Mertin, S ;
McDowall, SG ;
Harley, VR .
NUCLEIC ACIDS RESEARCH, 1999, 27 (05) :1359-1364
[7]   SEX-REVERSING MUTATIONS AFFECT THE ARCHITECTURE OF SRY-DNA COMPLEXES [J].
PONTIGGIA, A ;
RIMINI, R ;
HARLEY, VR ;
GOODFELLOW, PN ;
LOVELLBADGE, R ;
BIANCHI, ME .
EMBO JOURNAL, 1994, 13 (24) :6115-6124
[8]  
Schratter P, 1996, Methods Mol Biol, V59, P115
1