期刊
BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH
卷 1745, 期 2, 页码 254-264出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.bbamcr.2005.05.002
关键词
GoLoco; G-protein; GPSM2; LGN
资金
- NIGMS NIH HHS [GM065533, R01 GM062338] Funding Source: Medline
GPSM2 (G-protein signalling modulator 2; also known as LGN or mammalian Pins) is a protein that regulates mitotic spindle organization and cell division. GPSM2 contains seven tetratricopeptide repeats (TPR) and four G alpha(i/o)-Loco (GoLoco) motifs. GPSM2 has guanine nucleotide dissociation inhibitor (GDI) activity towards both G alpha(o)- and G alpha(i)-subunits; however, a systematic analysis of its individual GoLoco motifs has not been described. We analyzed each of the four individual GoLoco motifs from GPSM2, assessing their relative binding affinities and GDI potencies for G alpha(i1), G alpha(i2), and G alpha(i3) and G alpha(o). Each of the four GPSM2 GoLoco motifs (36-43 amino acids in length) was expressed in bacteria as a GST-fusion protein and purified to homogeneity. The binding of each of the four GST-GoLoco motifs to G alpha(i1)-, G alpha(o)-, and G alpha(s)-subunits was assessed by surface plasmon resonance; all of the motifs bound G alpha(iI), but exhibited low affinity towards G alpha(o). GDI activity was assessed by a fluorescence-based nucleotide-binding assay, revealing that all four GoLoco motifs are functional as GDIs for G alpha(i1), G alpha(i2), and G alpha(i3). Consistent with our binding studies, the GDI activity of GPSM2 GoLoco motifs on Ga-o was significantly lower than that toward G alpha(i1), suggesting that the in vivo targets of GPSM2 are most likely to be Gai-subunits. (c) 2005 Elsevier B.V. All rights reserved.
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