Evidence for an inhibitory LIM domain in a rat brain agmatinase-like protein

We recently cloned a rat brain agmatinase-like protein (ALP) whose amino acid sequence greatly differs from other agmatinases and exhibits a LIM-like domain close to its carboxyl terminus. The protein was immunohistochemically detected in the hypothalamic region and hippocampal astrocytes and neurons. We now show that truncated species, lacking the LIM-type domain, retains the dimeric structure of the wild-type protein but exhibits a 10-fold increased kcat, a 3-fold decreased Km value for agmatine and altered intrinsic tryptophan fluorescent properties. As expected for a LIM protein, zinc was detected only in the wild-type ALP (∼2 Zn2+/monomer). Our proposal is that the LIM domain functions as an autoinhibitory entity and that inhibition is reversed by interaction of the domain with some yet undefined brain protein.

► Agmatinase-like protein (ALP) is a rat brain enzyme that catalyzes agmatine hydrolysis. ► We analyzed the function of a LIM-domain sequence in this protein. ► The catalytic efficiency was increased by about 30 times by removal of the LIM domain in ALP. ► We propose the LIM sequence as an autoinhibitory domain in ALP.