Autolysis at the disintegrin domain of patagonfibrase, a metalloproteinase from Philodryas patagoniensis (Patagonia Green Racer; Dipsadidae) venom.
Patagonfibrase is a 57.5-kDa hemorrhagic metalloproteinase isolated from the venom of Philodryas patagoniensis (Patagonia Green Racer), a South American rear-fanged snake. Herein we demonstrate that patagonfibrase undergoes autolysis at its pH optimum (7.5) and at 37 degrees C, primarily producing a approximately 32.6 kDa fragment composed of disintegrin-like and cysteine-rich domains, as identified by mass spectrometry and N-terminal sequencing. The autolysis site for production of this fragment is similar to that observed for metalloproteinases from front-fanged Viperidae snake venoms. In the presence of Ca(2+), patagonfibrase was only partially autolysed, giving rise mainly to one fragment of approximately 52.2 kDa. In addition, calcium markedly enhanced the azocaseinolytic activity of patagonfibrase. Our findings contribute to the understanding of the structural and mechanistic bases of this family of metalloenzymes that are widely distributed among snake venoms, demonstrating that important post-translational modifications such as proteolysis can also contribute to the diversity and complexity of proteins found in rear-fanged snake venoms.
Peichoto
María E
ME
Cátedra de Farmacología, Facultad de Ciencias Veterinarias, Universidad Nacional del Nordeste, Sargento Cabral 2139, 3400, Corrientes, Argentina. mepeichoto@yahoo.com.ar
Paes Leme
Adriana F
AF
Pauletti
Bianca A
BA
Batista
Isabel Correia
IC
Mackessy
Stephen P
SP
Acosta
Ofelia
O
Santoro
Marcelo L
ML
D000595:Amino Acid Sequence; D001329:Autolysis; D002853:Chromatography, Liquid; D019483:Disintegrins / Q000378:metabolism*; D013058:Mass Spectrometry; D045726:Metalloproteases / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D012910:Snake Venoms / Q000737:chemistry / Q000201:enzymology* / Q000378:metabolism
D019483:Disintegrins; D012910:Snake Venoms; D045726:Metalloproteases; C519455:patagonfibrase, Philodryas patagoniensis