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PMID: 9355733 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Lysosomal alpha-mannosidases of mouse tissues: characteristics of the isoenzymes, and cloning and expression of a full-length cDNA.

The Biochemical journal ·Vol. 327 ( Pt 1) ·1997-10-01 ·Pages 45-9

Beccari T, Appolloni MG, Costanzi E, Stinchi S, Stirling JL, Della Fazia MA, Servillo G, Viola MP, Orlacchio A

Abstract

Lysosomal alpha-d-mannosidase from mouse tissues was separated into its constituent isoenzymes by DEAE-cellulose chromatography. Forms corresponding to the human isoenzymes B and A were present in testis, brain, spleen and kidney, whereas in epididymis and liver only the B form was present. Murine alpha-mannosidases A and B are glycoproteins and have pH optima, thermal stabilities and molecular masses similar to those of the human isoenzymes. A full-length cDNA (3.1 kb) containing the complete coding sequence for alpha-mannosidase was isolated from a mouse macrophage cDNA library. Comparison of the deduced amino acid sequences of human and mouse alpha-mannosidases showed that they had 75% identity and 83% similarity. Expression of this cDNA in COS cells showed that both the A and the B isoenzymes can arise from a single transcript. Northern blotting analysis showed a 10-fold range in the abundance of alpha-mannosidase mRNA in mouse tissues, with the highest levels found in epididymis, and the lowest in liver.

MeSH Terms
Amino Acid Sequence Animals Blotting, Northern COS Cells Chromatography, DEAE-Cellulose Chromosome Mapping Cloning, Molecular DNA, Complementary/genetics Gene Expression Humans Isoenzymes/chemistry,genetics,isolation & purification,metabolism Lysosomes/enzymology Mannosidases/chemistry,genetics,isolation & purification,metabolism Mice Molecular Sequence Data RNA, Messenger/genetics,metabolism Sequence Homology, Amino Acid alpha-Mannosidase
Chemicals
DNA, Complementary Isoenzymes RNA, Messenger Mannosidases alpha-Mannosidase
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Beccari T
Dipartimento di Biologia Cellulare e Molecolare, Università degli Studi di Perugia, Via del Giochetto, 06126 Perugia, Italy.
Appolloni M G
Costanzi E
Stinchi S
Stirling J L
Della Fazia M A
Servillo G
Viola M P
Orlacchio A
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj3270045
Published
1997-10-01
Pages
45-9
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1218761
Subset
IM
Databases
GENBANK
U29947
External Links
PubMed source DOI: 10.1042/bj3270045 PMC Full Text View journal details
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