title: Identification of carboxypeptidase N as an enzyme responsible for C-terminal cleavage of stromal cell-derived factor-1alpha in the circulation.
abstract: 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.
authors.affiliation: HIV and AIDS Malignancy Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. dd162w@nih.gov <dd162w@nih.gov>
authors.forename: David A
authors.initials: DA
authors.lastname: Davis
authors.forename: Kathleen E
authors.initials: KE
authors.lastname: Singer
authors.forename: Maria
authors.initials: M
authors.lastname: De La Luz Sierra
authors.forename: Masashi
authors.initials: M
authors.lastname: Narazaki
authors.forename: Fuquan
authors.initials: F
authors.lastname: Yang
authors.forename: Henry M
authors.initials: HM
authors.lastname: Fales
authors.forename: Robert
authors.initials: R
authors.lastname: Yarchoan
authors.forename: Giovanna
authors.initials: G
authors.lastname: Tosato
mesh_terms: 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
chemical_list: C515255:CXCL12 protein, human; D054377:Chemokine CXCL12; D018925:Chemokines; D019743:Chemokines, CXC; C515256:Cxcl12 protein, mouse; D011994:Recombinant Proteins; D001121:Lysine Carboxypeptidase; D008239:Lysine
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