Linked Life Data

10655158

Source:http://linkedlifedata.com/resource/pubmed/id/10655158

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
2000-2-29
pubmed:abstractText
Biotinidase deficiency is an autosomal recessive disorder of biotin recycling. Biotinidase cleaves the biotin from biocytin or short biotinyl-peptides to replenish the free biotin pool, or it can transfer the vitamin to specific proteins. The cDNA for human serum biotinidase has two in-frame start codons, potentially allowing for the synthesis of an enzyme with a signal peptide (SP) consisting of either 21 or 41 amino acids. In order to examine the requirements of the signal peptide region for the production and secretion of biotinidase, three different forms of the normal human serum biotinidase gene were constructed that encode either the 21-amino-acid SP (SP21-NL) or the 41-amino-acid SP (SP41-NL) or without a SP (NoSP-NL). These constructs were expressed in insect cells via a baculovirus expression system. Biotinidase from cells with SP41-NL and SP21-NL had immunoreactive and biotinyl-hydrolase-active enzyme in lysates and expression media. Cells with NoSP-NL had about 3% of the immunoreactive material and no enzyme activity in lysates and no immunoreactive protein or enzymatic activity in the expression medium. Lack of biotinidase from cells with NoSP-NL may be due to translation inefficiency or increased susceptibility of this species to protease degradation than the secreted forms. We have demonstrated that the 21-amino-acid signal peptide is sufficient to result in glycosylated, secreted biotinidase, but we cannot determine if the glycosylated biotinidase in the lysates or secreted in the medium of cells with SP41-NL use the first, second, or both ATGs in the SP region. Because this particular expression system has no mechanism for timing the movement of newly translated biotinidase protein, we cannot draw conclusions about the relative efficiency of SP41-NL versus SP21-NL, but it is possible that either is used in vivo depending on particular cellular conditions.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jan
pubmed:issn
1096-7192
pubmed:author
pubmed:copyrightInfo
Copyright 2000 Academic Press.
pubmed:issnType
Print
pubmed:volume
69
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
56-63
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed-meshheading:10655158-Amidohydrolases, pubmed-meshheading:10655158-Amino Acid Sequence, pubmed-meshheading:10655158-Animals, pubmed-meshheading:10655158-Baculoviridae, pubmed-meshheading:10655158-Biological Transport, pubmed-meshheading:10655158-Biotinidase, pubmed-meshheading:10655158-Cell Line, pubmed-meshheading:10655158-Codon, Initiator, pubmed-meshheading:10655158-Culture Media, Conditioned, pubmed-meshheading:10655158-Gene Expression, pubmed-meshheading:10655158-Glycosylation, pubmed-meshheading:10655158-Humans, pubmed-meshheading:10655158-Molecular Sequence Data, pubmed-meshheading:10655158-Molecular Weight, pubmed-meshheading:10655158-Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase, pubmed-meshheading:10655158-Protein Biosynthesis, pubmed-meshheading:10655158-Protein Sorting Signals, pubmed-meshheading:10655158-RNA, Messenger, pubmed-meshheading:10655158-Recombinant Proteins, pubmed-meshheading:10655158-Spodoptera
pubmed:year
2000
pubmed:articleTitle
Examination of the signal peptide region of human biotinidase using a baculovirus expression system.
pubmed:affiliation
Department of Human Genetics, Medical College of Virginia of Virginia Commonwealth University, Richmond, Virginia, 23298, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.