PAP I interacts with itself, PAP II, PAP III, and lithostathine/regIalpha

Abstract

PAP I, PAP II, PAP III, and lithostathine/regIalpha are members of a multigenic family of proteins expressed in several tissues. PAP I was shown to be antiapoptotic, mitogenic, and anti-inflammatory and can promote cell adhesion to the extracellular matrix. Lithostathine/regIalpha can be mitogenic. Because polymerization might regulate activity, we examined the ability of rat PAP I to interact with itself (homodimerization), PAP II, PAP III, and lithostathine/regIalpha (heterodimerization) by the yeast two-hybrid system, affinity experiments, and crosslinking. PAP I interacted significantly with all members of the PAP protein family, homodimerization showing the strongest interaction as judged by the beta-galactosidase test. This was confirmed by showing specific affinity between a MBP-rPAP I fusion protein and the native rPAP I. Finally, crosslinking experiments showed that rPAP I formed dimers in solution. These findings should be taken into account in functional studies involving PAP I and PAP-related proteins.