title: Location and stoichiometry of the protease CspB and the cortex-lytic enzyme SleC in Clostridium perfringens spores.
abstract: The protease CspB and the cortex-lytic enzyme SleC are essential for peptoglycan cortex hydrolysis during germination of spores of the Clostridium perfringens food poisoning isolate SM101. In this study, Western blot analyses were used to demonstrate that CspB and SleC are present exclusively in the C. perfringens SM101 spore coat layer fraction and absent in the lysate from decoated spores and from the purified inner spore membrane. These results indicate why decoating treatments greatly reduce both germination and apparent viability of C. perfringens spores in the absence of an exogenous lytic enzyme. In addition, quantitative Western blot analyses showed that there are approximately 2000 and 130,000 molecules of CspB and pro-SleC, respectively, per C. perfringens SM101 spore, consistent with CspB's role in acting catalytically on pro-SleC to convert this zymogen to the active enzyme.
authors.affiliation: Departments of Biomedical Sciences, Oregon State University, Corvallis, OR 97331, USA; Microbiology, Oregon State University, Corvallis, OR 97331, USA; Medical Laboratories Department, College of Science Al-Zulfi, Majmaah University, Saudi Arabia.
authors.forename: Saeed
authors.initials: S
authors.lastname: Banawas
authors.affiliation: Department of Molecular Biology and Biophysics, University of Connecticut Health Center, Farmington, CT 06030-3305, USA.
authors.forename: George
authors.initials: G
authors.lastname: Korza
authors.affiliation: Departments of Biomedical Sciences, Oregon State University, Corvallis, OR 97331, USA; Gut Microbiota and Clostridia Research Group, Departamento de Ciencias Biológicas, Facultad de Ciencias Biológicas, Universidad Andres Bello, Santiago, Chile.
authors.forename: Daniel
authors.initials: D
authors.lastname: Paredes-Sabja
authors.affiliation: Department of Molecular Biology and Biophysics, University of Connecticut Health Center, Farmington, CT 06030-3305, USA.
authors.forename: Yunfeng
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Molecular Biology and Biophysics, University of Connecticut Health Center, Farmington, CT 06030-3305, USA.
authors.forename: Bing
authors.initials: B
authors.lastname: Hao
authors.affiliation: Department of Molecular Biology and Biophysics, University of Connecticut Health Center, Farmington, CT 06030-3305, USA.
authors.forename: Peter
authors.initials: P
authors.lastname: Setlow
authors.affiliation: Departments of Biomedical Sciences, Oregon State University, Corvallis, OR 97331, USA; Microbiology, Oregon State University, Corvallis, OR 97331, USA. Electronic address: sarkerm@oregonstate.edu.
authors.forename: Mahfuzur R
authors.initials: MR
authors.lastname: Sarker
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D003016:Clostridium perfringens / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology; D006868:Hydrolysis; D013171:Spores, Bacterial / Q000201:enzymology* / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; C077188:cold-shock protein CspB, Bacteria
keywords: Clostridium perfringens; CspB; SleC; Spore germination; Spores