TY - JOUR
T1 - Tetraspanin protein CD9 interacts with metalloprotease CD10 and enhances its release via exosomes
AU - Mazurov, Dmitriy
AU - Barbashova, Lubov
AU - Filatov, Alexander
PY - 2013/3
Y1 - 2013/3
N2 - Tetraspanins interact with a wide variety of transmembrane and intracellular proteins called molecular partners, and modulate their function. In this article, we describe a new partner of tetraspanin web, membrane metalloprotease CD10, which is selectively associated with CD9. By constructing chimeras between tetraspanins CD9 and CD82 (the latter does not interact with CD10) or by using site-directed mutagenesis, we determined that a portion of the large extracellular loop from the CCG motif to transmembrane domain 4, as well as the C-terminal tail of CD9, are involved in the interaction with CD10. The stable expression of wild-type CD9 in K562 CD10-positive cells enhanced the level of CD10 released with exosomes five-fold. In contrast, the expression of chimeric CD9, which contained the cytoplasmic C-terminal domain from CD82, had little effect on CD10 release. Short hairpin RNA knockdown of CD9 expression in Nalm-6 pre-B cells resulted in a two-fold reduction in the amount of endogenous CD10 released with microvesicles. The peptidase activity of CD10 measured either on cells or on exosomes correlated with the level of CD10 expression, and was not significantly modulated by CD9 expression as such. Our data suggest that the interaction of CD10 with tetraspanin CD9 can play an important role in the redistribution of peptidase activity from the cell surface to outer microenvironments. In bone marrow, where CD10 presumably contributes to the maturation of pre-B cells and migration of B cells to the blood circulation, release of CD10 peptidase activity with exosomes may effectively regulate extracellular matrix microenvironments. Structured digital abstract CD9 physically interacts with CD179b, Tetraspanin-9, CD63, CD81, HLA class II, Uncharacterized protein C14orf166B, Tetraspanin-14, HLA class I, Galactokinase, Protein disulfide-isomerase A6, CD98, CD19, CD316, CD146, CD92, Annexin A6, CD156c, CD71, CD10, CD29, CD315, DAAM1 and CD49f by anti bait coimmunoprecipitation (View interaction) CD10 physically interacts with CD9 by anti bait coimmunoprecipitation (View Interaction: 1, 2, 3, 4) CD10 physically interacts with CD179b, Protein FAM207A, CD9, CD81, Uncharacterized protein C22orf13, HLA class II, Uncharacterized protein C3orf26, HLA class I, Protein C14orf21, Annexin A6 and DAAM1 by anti bait coimmunoprecipitation (View interaction) Tetraspanins interact with transmembrane proteins called molecular partners. We describe a new partner of tetraspanin CD9, metalloprotease CD10. Using mutagenic analysis we determine that large extracellular loop and cytoplasmic C-terminus of CD9 are involved in the interaction with CD10. Association of CD10 with CD9 enhances the release of CD10 with exosomes. This may facilitate pre-B cell migration in bone marrow
AB - Tetraspanins interact with a wide variety of transmembrane and intracellular proteins called molecular partners, and modulate their function. In this article, we describe a new partner of tetraspanin web, membrane metalloprotease CD10, which is selectively associated with CD9. By constructing chimeras between tetraspanins CD9 and CD82 (the latter does not interact with CD10) or by using site-directed mutagenesis, we determined that a portion of the large extracellular loop from the CCG motif to transmembrane domain 4, as well as the C-terminal tail of CD9, are involved in the interaction with CD10. The stable expression of wild-type CD9 in K562 CD10-positive cells enhanced the level of CD10 released with exosomes five-fold. In contrast, the expression of chimeric CD9, which contained the cytoplasmic C-terminal domain from CD82, had little effect on CD10 release. Short hairpin RNA knockdown of CD9 expression in Nalm-6 pre-B cells resulted in a two-fold reduction in the amount of endogenous CD10 released with microvesicles. The peptidase activity of CD10 measured either on cells or on exosomes correlated with the level of CD10 expression, and was not significantly modulated by CD9 expression as such. Our data suggest that the interaction of CD10 with tetraspanin CD9 can play an important role in the redistribution of peptidase activity from the cell surface to outer microenvironments. In bone marrow, where CD10 presumably contributes to the maturation of pre-B cells and migration of B cells to the blood circulation, release of CD10 peptidase activity with exosomes may effectively regulate extracellular matrix microenvironments. Structured digital abstract CD9 physically interacts with CD179b, Tetraspanin-9, CD63, CD81, HLA class II, Uncharacterized protein C14orf166B, Tetraspanin-14, HLA class I, Galactokinase, Protein disulfide-isomerase A6, CD98, CD19, CD316, CD146, CD92, Annexin A6, CD156c, CD71, CD10, CD29, CD315, DAAM1 and CD49f by anti bait coimmunoprecipitation (View interaction) CD10 physically interacts with CD9 by anti bait coimmunoprecipitation (View Interaction: 1, 2, 3, 4) CD10 physically interacts with CD179b, Protein FAM207A, CD9, CD81, Uncharacterized protein C22orf13, HLA class II, Uncharacterized protein C3orf26, HLA class I, Protein C14orf21, Annexin A6 and DAAM1 by anti bait coimmunoprecipitation (View interaction) Tetraspanins interact with transmembrane proteins called molecular partners. We describe a new partner of tetraspanin CD9, metalloprotease CD10. Using mutagenic analysis we determine that large extracellular loop and cytoplasmic C-terminus of CD9 are involved in the interaction with CD10. Association of CD10 with CD9 enhances the release of CD10 with exosomes. This may facilitate pre-B cell migration in bone marrow
KW - CD10
KW - CD9
KW - exosomes
KW - interaction
KW - tetraspanins
UR - http://www.scopus.com/inward/record.url?scp=84874460525&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84874460525&partnerID=8YFLogxK
U2 - 10.1111/febs.12110
DO - 10.1111/febs.12110
M3 - Article
C2 - 23289620
AN - SCOPUS:84874460525
SN - 1742-464X
VL - 280
SP - 1200
EP - 1213
JO - FEBS Journal
JF - FEBS Journal
IS - 5
ER -