Studies of membrane association of CLN3 protein.
The product of the CLN3 gene is a novel protein of unknown function. Simulations using amphiphacy algorithms have shown that structurally CLN3 may be another candidate for the family of membranous proteins. Signals controlling intracellular targeting of many membrane proteins are present as short sequences within their cytoplasmic domains. In fact, the sequence of CLN3 protein contains several such signaling sequences, which are conserved among mammals. First, at the N-terminus, potential N-myristoylation motif is present. Second, the C-terminal part of CLN3 protein contains both the dileucine motif, which is a potential lysosomal targeting signal, and the prenylation motif. There is scanty evidence of lysosomal and/or mitochondrial localization of CLN3 protein. However, the question of where the functional site of the cln3 protein exists in vivo remains unanswered. From theoretical calculations, we hypothesized that CLN3 should be an integral part of the membranous micro-environment. First, to test this hypothesis, we initiated detergent-partitioning experiments, localizing CLN3 predominantly in a pool of membranous protein. Further studies have shown that CLN3 protein integrates spontaneously with cellular membranes. Second, based on the prenylation results of CLN3 protein in vitro, we discussed the possible topological consequences of C-terminal fragment of CLN3 protein.
Kaczmarski
W
W
Department of Pathological Neurobiology, NYS Institute for Basic Research in Developmental Disabilities, Staten Island, New York 10314, USA.
Wisniewski
K E
KE
Golabek
A
A
Kaczmarski
A
A
Kida
E
E
Michalewski
M
M
D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism*; D002474:Cell-Free System; D016213:Cyclins; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008562:Membrane Glycoproteins / Q000378:metabolism*; D018832:Molecular Chaperones / Q000378:metabolism*; D017830:Octoxynol; D011092:Polyethylene Glycols / Q000378:metabolism; D017368:Protein Prenylation; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins
C079895:CLN3 protein, S cerevisiae; C105199:CLN3 protein, human; D016213:Cyclins; D008562:Membrane Glycoproteins; D018832:Molecular Chaperones; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40