Abstract
A post-proline cleaving enzyme [post-proline endopeptidase: EC 3.4.21.26] was purified from lamb brain by a series of column chromatographies on DEAE-Sephadex, hydroxyapatite and Sephadex G-150. The purified enzyme appeared homogeneous on disc gel and sodium dodecyl sulfate (SDS) gel electrophoreses. The enzyme was most active at pH 7.0 with carbobenzoxy-Gly-Pro-beta-naphthylamide (Z-Gly-Pro-2-NNap) as a substrate and catalyzed the hydrolysis of oxytocin, vasopressin, thyrotropin releasing hormone (TRH), substance P, luteinizing hormone releasing hormone (LH-RH), and angiotensin at the carboxyl side of their proline residues, except for the Pro2-Lys3 bond in substance P. From the results of subsite mapping using synthetic peptides, five subsites, S3 to S2', for substrate interaction with the enzyme were deduced to be present, and high stereospecificity was observed at S2, S1, and S1'. The isoelectric point of the enzyme was at pH 4.9, and the molecular weights estimated by gel filtration and SDS gel electrophoresis were 74,000 and 77,000, respectively. The enzyme was markedly inhibited by diisopropylphosphoro fluoridate (DFP), carbobenzoxy-Gly-Pro-chloromethyl ketone (Z-Gly-Pro-CH2Cl), p-chloromercuribenzoate (PCMB), Hg2+, and ...Continue Reading
Citations
Dec 1, 1990·Biological Chemistry Hoppe-Seyler·C SchönleinA Barth
Sep 1, 1986·Journal of Neurochemistry·E A Del BelA R Martins
Jul 1, 1983·Journal of Neurochemistry·S Wilk, M Orlowski
May 1, 1999·Development, Growth & Differentiation·K YoshidaM Morisawa
Mar 15, 1991·Experientia·T IidaW L Epstein
Mar 30, 2001·The Journal of Biological Chemistry·M N HarrisV J Harwood
Aug 14, 1999·The Journal of Biological Chemistry·A KimuraT Takahashi
Feb 15, 1986·Experientia·T KambaraS Shoji
Jul 11, 2000·Japanese Journal of Pharmacology·I OosukaK Yamanouchi
Apr 23, 1991·European Journal of Biochemistry·L Polgar
Aug 19, 2009·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Timo T MyöhänenPekka T Männistö
Dec 24, 2017·Biochemical and Biophysical Research Communications·Mohit WadhawanSushma Rathaur
Jan 12, 2010·Journal of Medicinal Chemistry·Janice LawandiNicolas Moitessier
Sep 29, 1998·Archives of Biochemistry and Biophysics·T KabashimaT Yoshimoto
Mar 14, 2002·Regulatory Peptides·Minh Tam LeGeorges Vauquelin
Apr 1, 1997·Insect Biochemistry and Molecular Biology·S OhtsukiS Natori
Jan 4, 2001·The International Journal of Biochemistry & Cell Biology·B M QuintoD E Casarini
Nov 1, 1996·Pharmacology, Biochemistry, and Behavior·Y ShishidoS Watanabe
Aug 18, 1995·Neuroscience Letters·N MiuraS Watanabe
May 23, 1998·The International Journal of Biochemistry & Cell Biology·D F Cunningham, B O'Connor
Jan 1, 1993·Life Sciences·W R WelchesC M Ferrario
Nov 26, 1991·European Journal of Pharmacology·J R AtackJ J Kulagowski
Nov 28, 1983·Life Sciences·S Wilk
Mar 1, 1990·Biochimica Et Biophysica Acta·N YamakawaA Nagamatsu
Apr 27, 1984·Biochimica Et Biophysica Acta·S MizutaniY Tomoda
Nov 1, 1994·Infection and Immunity·P L MäkinenS A Syed
Jan 1, 1985·The International Journal of Biochemistry·D J DalyR J Pennington
Sep 21, 1988·Biochimica Et Biophysica Acta·A MoriyamaM Sasaki
Aug 8, 1983·Neuroscience Letters·J P Burbach
Jul 20, 1992·Neuroscience Letters·T NakajimaT Ohe
Apr 1, 1991·Experimental and Molecular Pathology·T IidaW L Epstein
Aug 1, 1983·Archives of Biochemistry and Biophysics·G D Green, E Shaw
Oct 1, 1993·Journal of Dermatological Science·M KusuharaY Sasai
Oct 13, 2006·Clinica Chimica Acta; International Journal of Clinical Chemistry·Inger BrandtAnne-Marie Lambeir
Jan 2, 2007·Neuropeptides·J A García-HorsmanJ I Venäläinen
Aug 24, 2007·International Journal of Biological Macromolecules·Jae-Young JePyo-Jam Park