Identification of carboxypeptidase N as an enzyme responsible for C-terminal cleavage of stromal cell-derived factor-1alpha in the circulation.
The chemokine stromal-derived factor-1alpha (SDF-1alpha) is an essential regulator of hematopoiesis, lymphocyte homing, pre-B-cell growth, and angiogenesis. As SDF-1alpha is constitutively expressed in many tissues, chemokine function is mostly regulated by proteolytic degradation. Human serum cleaves the 68-amino acid chemokine, SDF-1alpha, at both termini. The enzyme or enzymes responsible for the removal of the carboxy-terminal lysine from SDF-1alpha, leading to significant reduction in biologic activity, have not been identified. Using a new biochemical assay for measuring the carboxy-terminal cleavage activity, we purified from serum and plasma a peptidase that specifically removes the carboxy-terminal lysine from SDF-1alpha and identified it as carboxypeptidase N (CPN, also known as kininase I, arginine carboxypeptidase, and anaphylotoxin inactivator). We demonstrate that SDF-1alpha in serum and plasma lacks the carboxy terminal lysine, and depletion of CPN from serum and plasma significantly reduces the SDF-1alpha carboxypeptidase activity. Purified CPN effectively and specifically removes the carboxy-terminal lysine from SDF-1alpha and significantly reduces the chemokine's biologic activity as a pre-B-cell growth factor and chemoattractant. Thus, in addition to its role as a regulator of the biologic activity of kinins and anaphylatoxins, CPN is an important regulator of the biologic activity of SDF-1alpha by reducing the chemokine-specific activity.
Davis
David A
DA
HIV and AIDS Malignancy Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. dd162w@nih.gov <dd162w@nih.gov>
Singer
Kathleen E
KE
De La Luz Sierra
Maria
M
Narazaki
Masashi
M
Yang
Fuquan
F
Fales
Henry M
HM
Yarchoan
Robert
R
Tosato
Giovanna
G
D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology; D015153:Blotting, Western; D045744:Cell Line, Tumor; D019070:Cell Lineage; D049109:Cell Proliferation; D054377:Chemokine CXCL12; D018925:Chemokines / Q000378:metabolism; D019743:Chemokines, CXC / Q000737:chemistry*; D002633:Chemotaxis; D002851:Chromatography, High Pressure Liquid; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D006410:Hematopoiesis; D006801:Humans; D015151:Immunoblotting; D047468:Immunoprecipitation; D008239:Lysine / Q000737:chemistry; D001121:Lysine Carboxypeptidase / Q000378:metabolism / Q000502:physiology*; D013058:Mass Spectrometry; D051379:Mice; D009389:Neovascularization, Pathologic; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013997:Time Factors
C515255:CXCL12 protein, human; D054377:Chemokine CXCL12; D018925:Chemokines; D019743:Chemokines, CXC; C515256:Cxcl12 protein, mouse; D011994:Recombinant Proteins; D001121:Lysine Carboxypeptidase; D008239:Lysine
J Lab Clin Med. 1987 Dec;110(6):798-806
3681119
Mol Pharmacol. 1994 Jan;45(1):1-8
8302267
Trends Pharmacol Sci. 1988 Aug;9(8):299-304
3074547
J Biol Chem. 1990 Jan 5;265(1):13-9
2378615
Science. 1993 Jul 30;261(5121):600-3
8342023
Proc Natl Acad Sci U S A. 1994 Mar 15;91(6):2305-9
8134392
Braz J Med Biol Res. 1994 Aug;27(8):1725-31
7749364
Genomics. 1995 Aug 10;28(3):495-500
7490086
Nature. 1996 Aug 15;382(6592):635-8
8757135
Nature. 1996 Aug 29;382(6594):829-33
8752280
J Immunol. 2000 Jul 15;165(2):1053-8
10878383
Eur J Neurosci. 2000 Jun;12(6):1857-66
10886327
Nature. 2001 Mar 1;410(6824):50-6
11242036
Eur J Immunol. 2001 Mar;31(3):699-707
11241273
J Immunol. 2001 May 15;166(10):6196-202
11342641
Nat Immunol. 2001 Jun;2(6):515-22
11376338
J Biol Chem. 2001 Aug 10;276(32):29839-45
11390394
Eur J Pharmacol. 2001 Oct 19;429(1-3):161-76
11698039
J Biol Chem. 2001 Nov 23;276(47):43503-8
11571304
Nat Med. 2001 Dec;7(12):1339-46
11726975
Blood. 2002 Apr 15;99(8):2703-11
11929756
J Biol Chem. 2002 May 3;277(18):15677-89
11867624
Nat Immunol. 2002 Jul;3(7):687-94
12068293
J Clin Invest. 2003 Jan;111(2):187-96
12531874
J Hum Genet. 2003;48(1):20-2
12560874
Immunity. 2004 Jun;20(6):707-18
15189736
J Clin Invest. 1970 Dec;49(12):2427-36
4098172
Ann Intern Med. 1980 Sep;93(3):443-5
7437116
Anal Biochem. 1981 Nov 15;118(1):113-9
7337205
Cardiovasc Res. 1984 Nov;18(11):690-6
6498875
J Clin Immunol. 1986 Jan;6(1):87-91
3958137
Prog Allergy. 1986;39:344-51
3562474
J Leukoc Biol. 2003 Nov;74(5):880-8
12960279
Blood. 2003 Dec 1;102(12):3900-5
12907452
Mol Immunol. 2004 Jan;40(11):785-93
14687935
Blood. 2004 Apr 1;103(7):2452-9
14525775
J Med Chem. 2004 Jun 3;47(12):3058-64
15163187
Nature. 1996 Aug 29;382(6594):833-5
8752281
J Exp Med. 1997 Jan 6;185(1):111-20
8996247
J Exp Med. 1996 Sep 1;184(3):1101-9
9064327
EMBO J. 1997 Dec 1;16(23):6996-7007
9384579
Science. 1998 Jan 16;279(5349):381-4
9430588
Blood. 1998 Jun 15;91(12):4523-30
9616148
Proc Natl Acad Sci U S A. 1998 Jun 9;95(12):6941-6
9618518
Nature. 1998 Jun 11;393(6685):591-4
9634237
Nature. 1998 Jun 11;393(6685):595-9
9634238
Proc Natl Acad Sci U S A. 1998 Aug 4;95(16):9448-53
9689100
Proc Natl Acad Sci U S A. 1998 Nov 24;95(24):14423-8
9826716
Science. 1999 Feb 5;283(5403):845-8
9933168
Blood. 1999 Mar 1;93(5):1612-21
10029590
Immunity. 1999 Apr;10(4):463-71
10229189
Am J Pathol. 1999 Apr;154(4):1125-35
10233851
J Biol Chem. 1999 Aug 20;274(34):23916-25
10446158
Biochem Pharmacol. 1962 Jul;11:585-92
13890520
Clin Chem. 1989 Jan;35(1):177
2910562