A urokinase receptor-associated protein with specific collagen binding properties

N Behrendt; O N Jensen; L H Engelholm; E Mørtz; M Mann; K Danø

A urokinase receptor-associated protein with specific collagen binding properties

N Behrendt, O N Jensen, L H Engelholm, E Mørtz, M Mann, K Danø

127 Citations (Scopus)

Abstract

The plasminogen activation cascade system, directed by urokinase and the urokinase receptor, plays a key role in extracellular proteolysis during tissue remodeling. To identify molecular interaction partners of these trigger proteins on the cell, we combined covalent protein cross-linking with mass spectrometry based methods for peptide mapping and primary structure analysis of electrophoretically isolated protein conjugates. A specific tri-molecular complex was observed upon addition of pro-urokinase to human U937 cells. This complex included the urokinase receptor, pro-urokinase, and an unknown, high molecular weight urokinase receptor-associated protein. The tryptic peptide mixture derived from a cross-linked complex of pro-urokinase and the latter protein was analyzed by nanoelectrospray tandem mass spectrometric sequencing. This analysis identified the novel protein as the human homologue of a murine membrane-bound lectin with hitherto unknown function. The human cDNA was cloned and sequenced. The protein, designated uPARAP, is a member of the macrophage mannose receptor protein family and contains a putative collagen-binding (fibronectin type II) domain in addition to 8 C-type carbohydrate recognition domains. It proved capable of binding strongly to a single type of collagen, collagen V. This collagen binding reaction at the exact site of plasminogen activation on the cell may lead to adhesive functions as well as a contribution to cellular degradation of collagen matrices.

Original language	English
Journal	The Journal of Biological Chemistry
Volume	275
Issue number	3
Pages (from-to)	1993-2002
Number of pages	10
ISSN	0021-9258
Publication status	Published - 21 Jan 2000

Keywords

Amino Acid Sequence
Animals
Cloning, Molecular
Collagen
Cross-Linking Reagents
Dose-Response Relationship, Drug
Glycosylation
Humans
Mannose-Binding Lectins
Mass Spectrometry
Membrane Glycoproteins
Mice
Molecular Sequence Data
Muscle, Smooth, Vascular
Plasminogen Activators
Protein Binding
Protein Structure, Tertiary
Receptors, Cell Surface
Receptors, Urokinase Plasminogen Activator
Sequence Homology, Amino Acid
U937 Cells
Journal Article
Research Support, Non-U.S. Gov't

Cite this

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title = "A urokinase receptor-associated protein with specific collagen binding properties",

abstract = "The plasminogen activation cascade system, directed by urokinase and the urokinase receptor, plays a key role in extracellular proteolysis during tissue remodeling. To identify molecular interaction partners of these trigger proteins on the cell, we combined covalent protein cross-linking with mass spectrometry based methods for peptide mapping and primary structure analysis of electrophoretically isolated protein conjugates. A specific tri-molecular complex was observed upon addition of pro-urokinase to human U937 cells. This complex included the urokinase receptor, pro-urokinase, and an unknown, high molecular weight urokinase receptor-associated protein. The tryptic peptide mixture derived from a cross-linked complex of pro-urokinase and the latter protein was analyzed by nanoelectrospray tandem mass spectrometric sequencing. This analysis identified the novel protein as the human homologue of a murine membrane-bound lectin with hitherto unknown function. The human cDNA was cloned and sequenced. The protein, designated uPARAP, is a member of the macrophage mannose receptor protein family and contains a putative collagen-binding (fibronectin type II) domain in addition to 8 C-type carbohydrate recognition domains. It proved capable of binding strongly to a single type of collagen, collagen V. This collagen binding reaction at the exact site of plasminogen activation on the cell may lead to adhesive functions as well as a contribution to cellular degradation of collagen matrices.",

keywords = "Amino Acid Sequence, Animals, Cloning, Molecular, Collagen, Cross-Linking Reagents, Dose-Response Relationship, Drug, Glycosylation, Humans, Mannose-Binding Lectins, Mass Spectrometry, Membrane Glycoproteins, Mice, Molecular Sequence Data, Muscle, Smooth, Vascular, Plasminogen Activators, Protein Binding, Protein Structure, Tertiary, Receptors, Cell Surface, Receptors, Urokinase Plasminogen Activator, Sequence Homology, Amino Acid, U937 Cells, Journal Article, Research Support, Non-U.S. Gov't",

author = "N Behrendt and Jensen, {O N} and Engelholm, {L H} and E M{\o}rtz and M Mann and K Dan{\o}",

year = "2000",

month = jan,

day = "21",

language = "English",

volume = "275",

pages = "1993--2002",

journal = "Journal of Biological Chemistry",

issn = "0021-9258",

publisher = "American Society for Biochemistry and Molecular Biology, Inc.",

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TY - JOUR

T1 - A urokinase receptor-associated protein with specific collagen binding properties

AU - Behrendt, N

AU - Jensen, O N

AU - Engelholm, L H

AU - Mørtz, E

AU - Mann, M

AU - Danø, K

PY - 2000/1/21

Y1 - 2000/1/21

N2 - The plasminogen activation cascade system, directed by urokinase and the urokinase receptor, plays a key role in extracellular proteolysis during tissue remodeling. To identify molecular interaction partners of these trigger proteins on the cell, we combined covalent protein cross-linking with mass spectrometry based methods for peptide mapping and primary structure analysis of electrophoretically isolated protein conjugates. A specific tri-molecular complex was observed upon addition of pro-urokinase to human U937 cells. This complex included the urokinase receptor, pro-urokinase, and an unknown, high molecular weight urokinase receptor-associated protein. The tryptic peptide mixture derived from a cross-linked complex of pro-urokinase and the latter protein was analyzed by nanoelectrospray tandem mass spectrometric sequencing. This analysis identified the novel protein as the human homologue of a murine membrane-bound lectin with hitherto unknown function. The human cDNA was cloned and sequenced. The protein, designated uPARAP, is a member of the macrophage mannose receptor protein family and contains a putative collagen-binding (fibronectin type II) domain in addition to 8 C-type carbohydrate recognition domains. It proved capable of binding strongly to a single type of collagen, collagen V. This collagen binding reaction at the exact site of plasminogen activation on the cell may lead to adhesive functions as well as a contribution to cellular degradation of collagen matrices.

AB - The plasminogen activation cascade system, directed by urokinase and the urokinase receptor, plays a key role in extracellular proteolysis during tissue remodeling. To identify molecular interaction partners of these trigger proteins on the cell, we combined covalent protein cross-linking with mass spectrometry based methods for peptide mapping and primary structure analysis of electrophoretically isolated protein conjugates. A specific tri-molecular complex was observed upon addition of pro-urokinase to human U937 cells. This complex included the urokinase receptor, pro-urokinase, and an unknown, high molecular weight urokinase receptor-associated protein. The tryptic peptide mixture derived from a cross-linked complex of pro-urokinase and the latter protein was analyzed by nanoelectrospray tandem mass spectrometric sequencing. This analysis identified the novel protein as the human homologue of a murine membrane-bound lectin with hitherto unknown function. The human cDNA was cloned and sequenced. The protein, designated uPARAP, is a member of the macrophage mannose receptor protein family and contains a putative collagen-binding (fibronectin type II) domain in addition to 8 C-type carbohydrate recognition domains. It proved capable of binding strongly to a single type of collagen, collagen V. This collagen binding reaction at the exact site of plasminogen activation on the cell may lead to adhesive functions as well as a contribution to cellular degradation of collagen matrices.

KW - Amino Acid Sequence

KW - Animals

KW - Cloning, Molecular

KW - Collagen

KW - Cross-Linking Reagents

KW - Dose-Response Relationship, Drug

KW - Glycosylation

KW - Humans

KW - Mannose-Binding Lectins

KW - Mass Spectrometry

KW - Membrane Glycoproteins

KW - Mice

KW - Molecular Sequence Data

KW - Muscle, Smooth, Vascular

KW - Plasminogen Activators

KW - Protein Binding

KW - Protein Structure, Tertiary

KW - Receptors, Cell Surface

KW - Receptors, Urokinase Plasminogen Activator

KW - Sequence Homology, Amino Acid

KW - U937 Cells

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

M3 - Journal article

C2 - 10636902

SN - 0021-9258

VL - 275

SP - 1993

EP - 2002

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

IS - 3

ER -

A urokinase receptor-associated protein with specific collagen binding properties

Abstract

Keywords

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