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Pepsin-mediated processing of the cytoplasmic histone H2A to strong antimicrobial peptide buforin I

 Hun Sik Kim  ;  Ho Yoon  ;  Il Minn  ;  Chan Bae Park  ;  Won Taek Lee  ;  Michael Zasloff  ;  Sun chang Kim 
 Journal of Immunology, Vol.165(6) : 3268-3274, 2000 
Journal Title
Issue Date
Amino Acid Sequence ; Animals ; Anti-Infective Agents/metabolism* ; Bufo bufo ; Cytoplasm/metabolism ; Gastric Mucosa/chemistry ; Gastric Mucosa/metabolism ; Gastric Mucosa/secretion ; Gastric Mucosa/ultrastructure ; Histones/metabolism* ; Hydrolysis ; Immunohistochemistry ; Microscopy, Immunoelectron ; Molecular Sequence Data ; Pepsin A/isolation & purification ; Pepsin A/physiology* ; Protein Biosynthesis ; Protein Precursors/metabolism ; Protein Processing, Post-Translational* ; Proteins/metabolism* ; Proteins/secretion ; Swine
The intestinal epithelium forms a first line of innate host defense by secretion of proteins with antimicrobial activity against microbial infection. Despite the extensive studies on the antimicrobial host defense in many gastrointestinal tracts, little is known about the antimicrobial defense system of the stomach. The potent antimicrobial peptide buforin I, consisting of 39 aa, was isolated recently from the stomach tissue of an Asian toad, Bufo bufo gargarizans. In this study we examined the mechanism of buforin I production in toad stomach tissue. Buforin I is produced by the action of pepsin isozymes, named pepsin Ca and Cb, cleaving the Tyr39-Ala40 bond of histone H2A. Immunohistochemical analysis revealed that buforin I is present extracellularly on the mucosal surface, and unacetylated histone H2A, a precursor of buforin I, is localized in the cytoplasm of gastric gland cells. Furthermore, Western blot analysis showed that buforin I is also present in the gastric fluids, and immunoelectron microscopy detected localization of the unacetylated histone H2A in the cytoplasmic granules of gastric gland cells. The distinct subcellular distribution of the unacetylated histone H2A and the detection of the unacetylated buforin I both on the mucosal surface and in the lumen suggest that buforin I is produced from the cytoplasmic unacetylated histone H2A secreted into the gastric lumen and subsequently processed by pepsins. Our results indicate that buforin I along with pepsins in the vertebrate stomach may contribute to the innate host defense of the stomach against invading microorganisms
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Yonsei Authors
Lee, Won Taek(이원택) ORCID logo https://orcid.org/0000-0001-7348-9562
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