Purification and characterization of novel heparin-binding growth factors in uterine secretory fluids - Identification as heparin-regulated M-r 10,000 forms of connective tissue growth factor

被引:180
作者
Brigstock, DR
Steffen, CL
Kim, GY
Vegunta, RK
Diehl, JR
Harding, PA
机构
[1] OHIO STATE UNIV, DEPT SURG, COLUMBUS, OH 43205 USA
[2] OHIO STATE UNIV, DEPT MED BIOCHEM, COLUMBUS, OH 43205 USA
[3] OHIO STATE UNIV, MOL CELLULAR & DEV BIOL PROGRAM, COLUMBUS, OH 43205 USA
[4] CLEMSON UNIV, DEPT ANIM DAIRY & VET SCI, CLEMSON, SC 29634 USA
关键词
D O I
10.1074/jbc.272.32.20275
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Uterine growth factors are potential effector molecules in embryo growth signaling pathways. Pig uterine luminal flushings contained a heparin-binding growth factor (HBGF) that required 0.8 M NaCl for elution from heparin columns and was termed HBGF-0.8, This factor, which was heat- and acid-labile and of M-r 10,000 as assessed by gel filtration, stimulated DNA synthesis in fibroblasts and smooth muscle cells but not endothelial cells. Two forms of HBGF-0.8, termed HBGF-0.8-P1 and HBGF-0.8-P2, exhibited differential heparin-binding properties. SDS-polyacrylamide gel electrophoresis showed that each form of HBGF-0.8 migrated with an apparent M-r of 10,000 under reducing conditions, Amino acid sequencing revealed the N-terminal sequence EENIKKGKKXIRTPKI for HBGF-0.8-P1 and EENIKKGKKXIRT for HBGF-0.8-P2. These sequences corresponded, respectively, to residues 247-262 and 248-259 of the 349-residue predicted primary translation product of porcine connective tissue growth factor (pCTGF). 10-kDa CTGF-mediated fibroblast DNA synthesis was modulated by exogenous heparin, and CTGF-immunoreactive proteins of 10, 16, and 20 kDa were present in unfractionated uterine luminal flushings. These data reveal the identity of a novel growth factor in uterine fluids as a highly truncated form of CTGF and show that the N-terminal two-thirds of the CTGF primary translation product is not required for mitogenic activity or heparin binding.
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页码:20275 / 20282
页数:8
相关论文
共 46 条
[1]   GROWTH, PROTEIN-CONTENT AND DISTRIBUTION OF EARLY PIG EMBRYOS [J].
ANDERSON, LL .
ANATOMICAL RECORD, 1978, 190 (01) :143-&
[2]   TEMPORAL EXPRESSION AND LOCATION OF COLONY-STIMULATING FACTOR-I (CSF-1) AND ITS RECEPTOR IN THE FEMALE REPRODUCTIVE-TRACT ARE CONSISTENT WITH CSF-1-REGULATED PLACENTAL DEVELOPMENT [J].
ARCECI, RJ ;
SHANAHAN, F ;
STANLEY, ER ;
POLLARD, JW .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1989, 86 (22) :8818-8822
[3]   HEPARIN-DEPENDENT BINDING AND AUTOPHOSPHORYLATION OF EPIDERMAL GROWTH-FACTOR (EGF) RECEPTOR BY HEPARIN-BINDING EGF-LIKE GROWTH-FACTOR BUT NOT BY EGF [J].
AVIEZER, D ;
YAYON, A .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1994, 91 (25) :12173-12177
[4]   RECEPTOR-BINDING AND HEPARIN-BINDING DOMAINS OF BASIC FIBROBLAST GROWTH-FACTOR [J].
BAIRD, A ;
SCHUBERT, D ;
LING, N ;
GUILLEMIN, R .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (07) :2324-2328
[5]   ISOLATION AND CHARACTERIZATION OF A MACROPHAGE-DERIVED HEPARIN-BINDING GROWTH-FACTOR [J].
BESNER, G ;
HIGASHIYAMA, S ;
KLAGSBRUN, M .
CELL REGULATION, 1990, 1 (11) :811-819
[6]   Interaction of Heparin-Binding EGF-Like Growth Factor (HB-EGF) with the Epidermal Growth Factor Receptor: Modulation by Heparin, Heparinase, or Synthetic Heparin-Binding HB-EGF Fragments [J].
Besner, Gail E. ;
Whelton, Diane ;
Crissman-Combs, Melissa A. ;
Steffen, Christy L. ;
Kim, Gregory Y. ;
Brigstock, David R. .
GROWTH FACTORS, 1992, 7 (04) :289-296
[7]   THE MODULAR ARCHITECTURE OF A NEW FAMILY OF GROWTH-REGULATORS RELATED TO CONNECTIVE-TISSUE GROWTH-FACTOR [J].
BORK, P .
FEBS LETTERS, 1993, 327 (02) :125-130
[8]   CONNECTIVE-TISSUE GROWTH-FACTOR - A CYSTEINE-RICH MITOGEN SECRETED BY HUMAN VASCULAR ENDOTHELIAL-CELLS IS RELATED TO THE SRC-INDUCED IMMEDIATE EARLY GENE-PRODUCT CEF-10 [J].
BRADHAM, DM ;
IGARASHI, A ;
POTTER, RL ;
GROTENDORST, GR .
JOURNAL OF CELL BIOLOGY, 1991, 114 (06) :1285-1294
[9]  
BRAY W, 1994, AM LAB, V26, P38
[10]   Pig uterine luminal fluid contains the developmentally regulated neurotrophic factor, pleiotrophin [J].
Brigstock, DR ;
Kim, GY ;
Steffen, CL .
JOURNAL OF ENDOCRINOLOGY, 1996, 148 (01) :103-111