Characterization of MARCKS (Myristoylated Alanine-Rich C Kinase Substrate) Identified by a Monoclonal Antibody Generated against Chick Embryo Neural Retina

Abstract

To identify molecular markers of cell differentiation in developing nervous tissue, monoclonal antibodies against chick embryo neural retina were made. One of them, 3C3mAb, recognized a developmentally regulated antigen present in several organs of the CNS. Data from MALDI-TOF mass spectrometry and peptide sequencing of the immuno-affinity purified protein indicated identity of the antigen with MARCKS. The immunoreactive material was always found as a unique polypeptide (Mr71 kDa) in SDS–PAGE, however isoelectrofocusing revealed the existence of several bands (pIranging from 4.0 to 4.5). Interestingly some retinal cell types, as photoreceptors, exhibited an extremely significant decrease in the intensity of the immunoreactive material during the final phases of terminal differentiation while others, as some retinal neurons, maintained the immunoreactivity when fully differentiated. Taken together these results indicate that MARCKS, a protein susceptible of several posttranslational modifications as myristoylation and phosphorylation at variable extent, may act differently in neural retina cell types.