Expression, purification and characterization of yellow grouper Epinephelus awoara regulator of G protein signaling 16 protein.
Regulators of G-protein signaling (RGS) proteins are a family of proteins, which accelerate GTPase-activity intrinsic to the alpha subunits of heterotrimeric G-proteins and play crucial roles in the physiological control of G-protein signaling. Here, yellow grouper RGS16 protein was expressed in Escherichia coli and purified by Ni-NTA affinity chromatography. The expression level of the fusion protein was up to 30% of the total cellular protein.Western blotting analysis showed that a band with the molecular mass of about 21 Kda was detected. The purified recombinant protein was used to prepare polyclonal antibody, and antiserum obtained was highly specific with the titer of over 1:32,000. Additionally, RGS16 protein was expressed in the Tn-5B1-4 insect cells. Western blotting analysis revealed that the expressed protein had immunoreactivity.
Key Laboratory of Animal Genetics and Breeding of State Ethnic Affairs Commission and Ministry of Education, College of Life Science and Technology, SouthWest University for Nationalities, 610041 Chengdu, China.
Li
L
Wang
Xinzhong
X
Wu
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation / Q000502:physiology*; D008969:Molecular Sequence Data; D009036:Moths / Q000166:cytology; D010473:Perciformes / Q000378:metabolism*; D010802:Phylogeny; D020710:RGS Proteins / Q000235:genetics / Q000378:metabolism*; D013154:Spleen / Q000378:metabolism
D020710:RGS Proteins; C107413:RGS16 protein
Cell. 1996 Jan 12;84(1):115-25
8548815
Biochem J. 2000 Dec 15;352 Pt 3:747-53
11104682
Postepy Hig Med Dosw (Online). 2004;58:312-20
15459549
Semin Cell Dev Biol. 2006 Jun;17(3):363-76
16687250
J Biol Chem. 1997 Feb 14;272(7):3871-4
9064301
J Neurochem. 2005 Sep;94(5):1212-24
16045448
Biochem Biophys Res Commun. 1999 Jun 16;259(3):550-6
10364456
Immunity. 1998 Jan;8(1):1-10
9462506
J Immunol. 1993 May 1;150(9):3895-904
8473738
Annu Rev Biochem. 2000;69:795-827
10966476
FEBS Lett. 1995 Mar 27;362(1):1-4
7535249
Nucleic Acids Res. 1994 Nov 11;22(22):4673-80
7984417
Proc Natl Acad Sci U S A. 1997 Jul 22;94(15):7868-72
9223279
Biochim Biophys Acta. 2002 Jan 30;1542(1-3):95-105
11853883
J Biol Chem. 2003 May 23;278(21):19301-8
12642593
Arch Immunol Ther Exp (Warsz). 2004 Jan-Feb;52(1):27-35
15053230
Int J Biol Sci. 2005;1(2):51-66
15951850
J Immunol. 1999 Mar 1;162(5):2677-82
10072511
Anal Biochem. 1976 May 7;72:248-54
942051
J Biol Chem. 1998 Jan 16;273(3):1269-72
9430654
Cell Mol Life Sci. 2006 Sep;63(17):1969-85
16847579
Annu Rev Pharmacol Toxicol. 2000;40:235-71
10836135
DNA Cell Biol. 1994 Feb;13(2):125-47
8179820
Mol Biol Evol. 1987 Jul;4(4):406-25
3447015
Brief Bioinform. 2004 Jun;5(2):150-63
15260895
Mol Cell Biol. 1996 Sep;16(9):5194-209
8756677
Nature. 1996 Sep 12;383(6596):175-7
8774883