Partial characterization of an approximately 20 K M(r) retinal protein whose phosphorylation is inhibited by taurine.
It has been demonstrated previously that taurine (2-aminoethanesulfonic acid) is an inhibitor of protein phosphorylation in a mitochondrial fraction of the rat retina. It appears that taurine is most effective in inhibiting the phosphorylation of an approximately 20 K apparent molecular weight (M(r)) protein found in the retinal tissue. This study further characterizes the location of the approximately 20 K phosphoprotein by phase separation using Triton X-114 and also characterizes the nature of the phosphate bond by various solvent extractions and by exposure to acid and base conditions. Triton X-114 experiments indicated that the approximately 20K phosphoprotein is located in the aqueous phase and, consequently, is probably not an integral protein of the mitochondrial membranes. Treatment of the phosphoprotein with solvents, acid, and/or base determined the phosphate linkage to be through a phosphoester bond rather than an acylphosphate bond. The approximately 20 K M(r) phosphoprotein was also isolated from one-dimensional polyacrylamide gels and subsequently digested with trypsin and hydrolyzed with 1 M HCl to break all peptide bonds. Analysis of the phosphoamino acids by two-dimensional high voltage electrophoresis on cellulose plates revealed that it is both the serine and threonine residues that are phosphorylated. However, phosphorylation of the serine residue(s) is predominant.
Lombardini
J B
JB
Department of Pharmacology, Texas Tech University Health Sciences Center, Lubbock 79430.
D000818:Animals; D004305:Dose-Response Relationship, Drug; D005136:Eye Proteins / Q000737:chemistry* / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism; D010750:Phosphoproteins / Q000737:chemistry*; D010766:Phosphorylation / Q000187:drug effects; D051381:Rats; D017208:Rats, Wistar; D012160:Retina / Q000378:metabolism*; D012694:Serine / Q000737:chemistry; D013654:Taurine / Q000494:pharmacology*; D013912:Threonine / Q000737:chemistry
D005136:Eye Proteins; D010750:Phosphoproteins; D013654:Taurine; D013912:Threonine; D012694:Serine