Switch to
| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
3
|
| pubmed:dateCreated |
1997-7-7
|
| pubmed:databankReference | |
| pubmed:abstractText |
As part of our ongoing efforts to characterize the N-glycosylation pathway of lepidopteran insect cells, we have isolated an alpha 1,2-mannosidase homolog from an Sf9 cDNA library. This cDNA contains an open reading frame which encodes a 670 amino acid protein with a calculated molecular weight of 75,225 Da. This protein has two potential N-glycosylation sites, two consensus calcium binding sequences, and is predicted to be a type II integral membrane protein with a 22 amino acid transmembrane domain (residues 31-52). The amino acid sequence of this protein is 35-57% identical to Drosophila, human, murine, and yeast alpha 1,2-mannosidases. A transcript of approximately 6 kilobases was detected by Northern blot analysis of Sf9 mRNA. Genomic Southern blots probed with an intron-free fragment of the alpha 1,2-mannosidase gene indicated that there are at least two copies or cross-hybridizing variants of this gene in the Sf9 genome. In vivo expression of the cDNA using a recombinant baculovirus produced a protein that released [3H]mannose from [3H]Man9GlcNAc. This activity required calcium, but not magnesium, and was inhibited by 1-deoxymannojirimycin. These results indicate that Sf9 cells encode and express an alpha 1,2-mannosidase with properties similar to those of other eukaryotic processing alpha 1,2-mannosidases.
|
| pubmed:grant | |
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical | |
| pubmed:status |
MEDLINE
|
| pubmed:month |
Apr
|
| pubmed:issn |
0959-6658
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:volume |
7
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
433-43
|
| pubmed:dateRevised |
2007-11-14
|
| pubmed:meshHeading |
pubmed-meshheading:9147053-Amino Acid Sequence,
pubmed-meshheading:9147053-Animals,
pubmed-meshheading:9147053-Baculoviridae,
pubmed-meshheading:9147053-Base Sequence,
pubmed-meshheading:9147053-Blotting, Southern,
pubmed-meshheading:9147053-Cell Line,
pubmed-meshheading:9147053-DNA, Complementary,
pubmed-meshheading:9147053-Drosophila,
pubmed-meshheading:9147053-Gene Expression,
pubmed-meshheading:9147053-Glycosylation,
pubmed-meshheading:9147053-Humans,
pubmed-meshheading:9147053-Mannosidases,
pubmed-meshheading:9147053-Mice,
pubmed-meshheading:9147053-Molecular Sequence Data,
pubmed-meshheading:9147053-Polymerase Chain Reaction,
pubmed-meshheading:9147053-Saccharomyces cerevisiae,
pubmed-meshheading:9147053-Sequence Homology,
pubmed-meshheading:9147053-Spodoptera,
pubmed-meshheading:9147053-Transfection
|
| pubmed:year |
1997
|
| pubmed:articleTitle |
Isolation and characterization of an alpha 1,2-mannosidase cDNA from the lepidopteran insect cell line Sf9.
|
| pubmed:affiliation |
Department of Entomology, Texas A&M University, College Station 77843, USA.
|
| pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.
|