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Query: EC:3.5.1.52 (
PNGase F
)
1,527
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
Murine
interleukin 5
(
IL-5
), a lymphokine produced by helper T cells, is involved in the regulation of growth and differentiation of B cells and other hematopoietic cells. The receptor for
IL-5
has been identified as two cross-linked complexes on T88-M cells (a murine
IL-5
-dependent early B cell line). In this study the
IL-5
receptor was directly characterized by utilizing an immobilized
IL-5
column and a rat monoclonal antibody, designated H7, directed against the
IL-5
receptor. H7 completely inhibited specific binding of 35S-labeled
IL-5
to T88-M cells, and bound to
IL-5
-responsive cells, e.g. T88-M, BCL1-B20 (a chronic B-cell leukemia), and MOPC104E (a myeloma), whereas H7 did not bind to
IL-5
-non-responsive cells, e.g. X5563 (a myeloma), FDC-P1 (an IL-3-dependent line), and MTH (an IL-2-dependent CTLL). H7 could barely bind to T88-M cells in the presence of
IL-5
, and immunoprecipitated a major band with an Mr of approximately 60 kd from the extract of surface-radioiodinated T88-M cells. The precipitation of this 60 kd molecule was inhibited by the addition of
IL-5
. Analysis with immobilized
IL-5
also revealed that a 60 kd molecule bound specifically to
IL-5
-coupled beads compared with control beads. Furthermore, no additional molecule with a higher Mr that was recognized by H7 appeared under non-reducing, compared with reducing, conditions. The 60 kd molecule recognized by H7 could be digested with
N-glycanase
to yield a protein band of approximately 55 kd.(ABSTRACT TRUNCATED AT 250 WORDS)
...
PMID:Characterization of the murine interleukin 5 receptor by using a monoclonal antibody. 208 84
IL-5
is a T cell-derived lymphokine that induces B cell growth and differentiation in murine systems. In this study, we examined the role of carbohydrate moiety of
IL-5
in the expression of biological function.
IL-5
polypeptides translated in Xenopus oocytes were heterogeneous in terms of isoelectric point (pI 4.7 to 8.0) and m.w. (45,000 to 60,000 under nonreducing conditions) and yielded m.w. of 25,000 to 30,000 under reducing conditions. Treatment of rIL-5 with
N-glycanase
under reducing conditions yielded an
IL-5
monomer of m.w. 12,000 to 14,000. Furthermore, deglycosylated rIL-5 that had been translated in the presence of tunicamycin showed very limited heterogeneity by two-dimensional gel electrophoresis (first dimension, nonequilibrium pH gradient electrophoresis; second dimension, SDS-PAGE). The m.w. was 27,000 to 28,000 under non-reducing conditions and migrated to m.w. 13,000 to 14,000 under reducing conditions. These results indicate that
IL-5
is a glycoprotein carrying the N-glycosidically-linked carbohydrates. Treatment of
IL-5
with sialidase caused the decrease in the heterogeneity in isoelectric point of
IL-5
. Deglycosylated rIL-5 that had been obtained from tunicamycin-treated oocytes could bind to
IL-5
-responding cells (T88-M), which express both high- and low-affinity
IL-5
receptors, as efficient as intact rIL-5 under high-affinity conditions. Scatchard plot analysis of equilibrium binding of 35S-labeled rIL-5 to T88-M cells revealed that the dissociation constants (Kd) of glycosylated rIL-5 and deglycosylated rIL-5 were 127 pM and 110 pM, respectively.
IL-5
activities determined by both B cell growth and differentiation assays were not affected by deglycosylation. These results indicate that N-linked glycoside moiety of
IL-5
molecules may not play an essential role in the expression of its activity.
...
PMID:Role of carbohydrate moiety of IL-5. Effect of tunicamycin on the glycosylation of IL-5 and the biologic activity of deglycosylated IL-5. 230 8
