Molecular cloning of the sphingolipid activator protein-1 (SAP-1), the sulfatide sulfatase activator.
A cDNA coding for SAP-1 was isolated from a lambda gt11 human hepatoma expression library using polyclonal antibodies raised against human SAP-1. Three positive clones were isolated with inserts of approximately 0.3 Kb (S1.1), 2 Kb (S1.2) and 2.2 Kb (S-1.3). The latter 2 contained an internal EcoRI site. All three clones cross-hybridized with one another, indicating sequence homology. The nucleotide sequence of S-1.1 was determined. Colinearity was established between 19 amino acids obtained by sequencing the amino terminus of pure SAP-1 and 57 bp from the 5' end of S-1.1. The open reading frame of S-1.1 coded for 67 amino acids. One glycosylation site was found 21 residues from the amino terminus, and no stop codons were found. S-1.1 codes for a mature polypeptide chain with a calculated molecular weight of 8955 daltons, corresponding to approximately 99% of mature SAP-1.
Dewji
N
N
Wenger
D
D
Fujibayashi
S
S
Donoviel
M
M
Esch
F
F
Hill
F
F
O'Brien
J S
JS
D000595:Amino Acid Sequence; D001483:Base Sequence; D002553:Cerebroside-Sulfatase / Q000378:metabolism*; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D006023:Glycoproteins; D006801:Humans; D009693:Nucleic Acid Hybridization; D011506:Proteins / Q000235:genetics*; D049231:Saposins; D049128:Sphingolipid Activator Proteins; D013429:Sulfatases / Q000378:metabolism*
D006023:Glycoproteins; C483345:PSAP protein, human; D011506:Proteins; D049231:Saposins; D049128:Sphingolipid Activator Proteins; D004247:DNA; D013429:Sulfatases; D002553:Cerebroside-Sulfatase