Structure-function analysis of GNIP, the glycogenin-interacting protein.
Glycogenin is a self-glucosylating protein that initiates glycogen biosynthesis. We recently identified a family of proteins, GNIPs, that interact with glycogenin and stimulate its self-glucosylating activity [J. Biol. Chem. 277 (2002) 19331]. The GNIP gene (also called TRIM7) encodes at least four distinct isoforms of GNIP, three of which (GNIP1, GNIP2, and GNIP3) have in common a COOH-terminal B30.2 domain and predicted coiled-coil regions. Based on Western blot analysis, the GNIP1 protein is widely distributed in tissues. From analysis of a series of deletion mutants of GNIP2 using the yeast two-hybrid system, the B30.2 domain was found to be responsible for the interaction with glycogenin. A truncated form of recombinant GNIP2, lacking the NH2-terminal coiled-coil region, was cross-linked to glycogenin by glutaraldehyde treatment, supporting the idea that the B30.2 domain was sufficient for the interaction. In the course of this study, GNIP2 was also found to interact with itself, via the coiled-coil domain. Heterologous interactions between GNIP1 and GNIP2 were also detected. Since glycogenin is also a dimer, higher order multimeric complexes between glycogenin and GNIPs would be possible.
Department of Biochemistry and Molecular Biology, Center for Diabetes Research, Indiana University School of Medicine, 635 Barnhill Drive, Indianapolis, IN 46202-5122, USA.
Lanmin
L
Zhai
Amy
A
Dietrich
Alexander V
AV
Skurat
Peter J
PJ
Roach
D000818:Animals; D015152:Blotting, Northern; D019556:COS Cells; D002352:Carrier Proteins / Q000737:chemistry* / Q000235:genetics / Q000502:physiology*; D005964:Glucosyltransferases; D006023:Glycoproteins / Q000378:metabolism; D051379:Mice; D009928:Organ Specificity; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000378:metabolism; D013329:Structure-Activity Relationship; D000070584:Tripartite Motif Proteins; D044767:Ubiquitin-Protein Ligases
D002352:Carrier Proteins; D006023:Glycoproteins; D012333:RNA, Messenger; D000070584:Tripartite Motif Proteins; C051887:glycogenin; C458504:TRIM7 protein, human; C482978:Trim7 protein, mouse; D044767:Ubiquitin-Protein Ligases; D005964:Glucosyltransferases