The noncatalytic portion of human UDP-glucose: glycoprotein glucosyltransferase I confers UDP-glucose binding and transferase function to the catalytic domain.

The eukaryotic cell monitors the fidelity of protein folding in the endoplasmic reticulum and only permits properly folded and/or assembled proteins to transit to the Golgi compartment in a process termed "quality control." An endoplasmic reticulum (ER) lumenal sensor for quality control is the UDP-glucose:glycoprotein glucosyltransferase that targets unfolded glycoproteins for transient, calcium-dependent glucosylation. This modification mediates glycoprotein interaction with the folding machinery comprised of calnexin or calreticulin in conjunction with ERp57. Two human UGT homologues, HUGT1 and HUGT2, exist that share 55% identity. The highest degree of identity resides in the COOH-terminal 20% of these proteins, the putative catalytic domain of HUGT1. However, only HUGT1 displays the expected functional activity. The contribution of the NH2-terminal remainder of HUGT1 to glucosyltransferase function is presently unknown. In this report we demonstrate that HUGT2 is localized to the ER in a manner that overlaps the distribution of HUGT1. Analysis of a series of HUGT1 and HUGT2 chimeric proteins demonstrated that the carboxyl-terminal region of HUGT2 contains a catalytic domain that is functional in place of the analogous portion of HUGT1. Whereas neither catalytic domain displayed detectable activity when expressed alone, co-expression of either catalytic domain with the noncatalytic amino-terminal portion of HUGT1 conferred UDP-Glc binding and transfer of glucose that was specific for unfolded glycoprotein substrates. The results indicate that the amino-terminal 80% of HUGT1 is required for activation of the catalytic domain, whereas the homologous portion of HUGT2 cannot provide this function.