Conformational changes in CD45 upon monoclonal antibody crosslinking

D Hamann; W Eichler; H Fiebig; R A van Lier

doi:10.1089/hyb.1996.15.11

Conformational changes in CD45 upon monoclonal antibody crosslinking

D Hamann, W Eichler, H Fiebig, R A van Lier

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

The CD45 protein tyrosine phosphatase is expressed in different isoforms that result from alternative splicing of three exons (A, B, and C) encoding regions near the N-terminus of the extracellular part of the molecule. We describe here a novel epitope on the N-terminal end of CD45 that is recognized by the MAb BL-TSub/2. Crossblocking studies showed that BL-TSub/2 and UCHL1 (CD45RO) binding sites are partially overlapping. However, in marked contrast to the CD45RO epitope, protease treatment of cells strongly diminished BL-TSub/2 binding. Similar to the UCHL1 epitope, the BL-TSub/2 binding site involves carbohydrate moieties, since neuraminidase treatment abrogated the reactivity of the MAb. Markedly, preincubation of cells with both CD45 common and CD45RA MAb induced a pronounced increase of BL-TSub/2 binding. This latter finding suggests that crosslinking of the CD45 molecule leads to conformational changes that could influence association of the molecule with putative ligands.

Original language	English
Pages (from-to)	11-6
Number of pages	6
Journal	Hybridoma
Volume	15
Issue number	1
DOIs	https://doi.org/10.1089/hyb.1996.15.11
Publication status	Published - Feb 1996
Externally published	Yes

Keywords

Animals
Antibodies, Monoclonal/pharmacology
Binding Sites, Antibody
Cross-Linking Reagents
Humans
Leukocyte Common Antigens/chemistry
Mice
Mice, Inbred BALB C

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10.1089/hyb.1996.15.11

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title = "Conformational changes in CD45 upon monoclonal antibody crosslinking",

abstract = "The CD45 protein tyrosine phosphatase is expressed in different isoforms that result from alternative splicing of three exons (A, B, and C) encoding regions near the N-terminus of the extracellular part of the molecule. We describe here a novel epitope on the N-terminal end of CD45 that is recognized by the MAb BL-TSub/2. Crossblocking studies showed that BL-TSub/2 and UCHL1 (CD45RO) binding sites are partially overlapping. However, in marked contrast to the CD45RO epitope, protease treatment of cells strongly diminished BL-TSub/2 binding. Similar to the UCHL1 epitope, the BL-TSub/2 binding site involves carbohydrate moieties, since neuraminidase treatment abrogated the reactivity of the MAb. Markedly, preincubation of cells with both CD45 common and CD45RA MAb induced a pronounced increase of BL-TSub/2 binding. This latter finding suggests that crosslinking of the CD45 molecule leads to conformational changes that could influence association of the molecule with putative ligands.",

keywords = "Animals, Antibodies, Monoclonal/pharmacology, Binding Sites, Antibody, Cross-Linking Reagents, Humans, Leukocyte Common Antigens/chemistry, Mice, Mice, Inbred BALB C",

author = "D Hamann and W Eichler and H Fiebig and {van Lier}, {R A}",

year = "1996",

month = feb,

doi = "10.1089/hyb.1996.15.11",

language = "English",

volume = "15",

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}

TY - JOUR

T1 - Conformational changes in CD45 upon monoclonal antibody crosslinking

AU - Hamann, D

AU - Eichler, W

AU - Fiebig, H

AU - van Lier, R A

PY - 1996/2

Y1 - 1996/2

N2 - The CD45 protein tyrosine phosphatase is expressed in different isoforms that result from alternative splicing of three exons (A, B, and C) encoding regions near the N-terminus of the extracellular part of the molecule. We describe here a novel epitope on the N-terminal end of CD45 that is recognized by the MAb BL-TSub/2. Crossblocking studies showed that BL-TSub/2 and UCHL1 (CD45RO) binding sites are partially overlapping. However, in marked contrast to the CD45RO epitope, protease treatment of cells strongly diminished BL-TSub/2 binding. Similar to the UCHL1 epitope, the BL-TSub/2 binding site involves carbohydrate moieties, since neuraminidase treatment abrogated the reactivity of the MAb. Markedly, preincubation of cells with both CD45 common and CD45RA MAb induced a pronounced increase of BL-TSub/2 binding. This latter finding suggests that crosslinking of the CD45 molecule leads to conformational changes that could influence association of the molecule with putative ligands.

AB - The CD45 protein tyrosine phosphatase is expressed in different isoforms that result from alternative splicing of three exons (A, B, and C) encoding regions near the N-terminus of the extracellular part of the molecule. We describe here a novel epitope on the N-terminal end of CD45 that is recognized by the MAb BL-TSub/2. Crossblocking studies showed that BL-TSub/2 and UCHL1 (CD45RO) binding sites are partially overlapping. However, in marked contrast to the CD45RO epitope, protease treatment of cells strongly diminished BL-TSub/2 binding. Similar to the UCHL1 epitope, the BL-TSub/2 binding site involves carbohydrate moieties, since neuraminidase treatment abrogated the reactivity of the MAb. Markedly, preincubation of cells with both CD45 common and CD45RA MAb induced a pronounced increase of BL-TSub/2 binding. This latter finding suggests that crosslinking of the CD45 molecule leads to conformational changes that could influence association of the molecule with putative ligands.

KW - Animals

KW - Antibodies, Monoclonal/pharmacology

KW - Binding Sites, Antibody

KW - Cross-Linking Reagents

KW - Humans

KW - Leukocyte Common Antigens/chemistry

KW - Mice

KW - Mice, Inbred BALB C

U2 - 10.1089/hyb.1996.15.11

DO - 10.1089/hyb.1996.15.11

M3 - Article

C2 - 9064281

SN - 0272-457X

VL - 15

SP - 11

EP - 16

JO - Hybridoma

JF - Hybridoma

IS - 1

ER -

Conformational changes in CD45 upon monoclonal antibody crosslinking

Abstract

Keywords

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