Untitled

Payie, Kenneth G.; Weadge, Joel T.; Tanaka, Takuji; Yada, Rickey Y.

doi:10.1023/a:1005656208075

Cited by 4 publications

(3 citation statements)

References 13 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…An aspartic peptidase was identified in rice by Asakura et al, 84 purified, 85 and subsequently expressed in E. coli. 86 Payie et al 87 purified a new aspartic proteinase from leaves of Medicago sativa L. (alfalfa) 3320-fold. They also demonstrated inhibition by pepstatin A and cleavage of synthetic substrates containing -Phe*Phebonds.…”

Section: F Plant Aspartic Peptidasesmentioning

confidence: 99%

Structure and Mechanism of the Pepsin-Like Family of Aspartic Peptidases

2002

View full text Add to dashboard Cite

Section: F Plant Aspartic Peptidasesmentioning

confidence: 99%

Structure and Mechanism of the Pepsin-Like Family of Aspartic Peptidases

2002

View full text Add to dashboard Cite

“…They are found in all higher organisms, and their respective roles are well established, although structural and functional characteristics of APs in plants are least understood. Of practical interest among plant APs are their roles in plant pathogen resistance (3) as well as in senescence and postharvest physiology (4,5). Plant APs share the common AP bilobal structure; however, some contain an additional sequence of ϳ100 residues inserted within the C-terminal primary structure.…”

mentioning

confidence: 99%

Structure and Mechanism of the Saposin-like Domain of a Plant Aspartic Protease

Bryksa

Bhaumik

Magracheva

et al. 2011

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Many plant aspartic proteases contain an additional sequence of ϳ100 amino acids termed the plant-specific insert, which is involved in host defense and vacuolar targeting. Similar to all saposin-like proteins, the plant-specific insert functions via protein-membrane interactions; however, the structural basis for such interactions has not been studied, and the nature of plantspecific insert-mediated membrane disruption has not been characterized. In the present study, the crystal structure of the saposin-like domain of potato aspartic protease was resolved at a resolution of 1.9 Å , revealing an open V-shaped configuration similar to the open structure of human saposin C. Notably, vesicle disruption activity followed Michaelis-Menten-like kinetics, a finding not previously reported for saposin-like proteins including plant-specific inserts. Circular dichroism data suggested that secondary structure was pH-dependent in a fashion similar to influenza A hemagglutinin fusion peptide. Membrane effects characterized by atomic force microscopy and light scattering indicated bilayer solubilization as well as fusogenic activity. Taken together, the present study is the first report to elucidate the membrane interaction mechanism of plant saposin-like domains whereby pH-dependent membrane interactions resulted in bilayer fusogenic activity that probably arose from a viral type pH-dependent helix-kink-helix motif at the plant-specific insert N terminus. Aspartic proteases (APs)2 are characterized by a common bilobal tertiary structure containing two catalytic aspartic acid residues (Asp 32 and Asp 215 in pepsin) within an active site cleft(1, 2). They are found in all higher organisms, and their respective roles are well established, although structural and functional characteristics of APs in plants are least understood. Of practical interest among plant APs are their roles in plant pathogen resistance (3) as well as in senescence and postharvest physiology (4, 5). Plant APs share the common AP bilobal structure; however, some contain an additional sequence of ϳ100 residues inserted within the C-terminal primary structure. These additional amino acids unique to plant APs (6 -8) create an extra domain protruding from the canonical AP molecule (9 -11). This structural oddity among APs is called the plantspecific insert (PSI), also known as the plant-specific sequence, which belongs to the saposin-like protein (SAPLIP) family (12, 13). Plant APs are found in either monomeric or heterodimeric forms (9, 14); the latter result from post-translational proteolysis, which includes the removal of part or all of the PSI, whereas the PSI is retained in monomeric plant APs (6,8).In general, members of the SAPLIP family have various physiological functions, all of which entail membrane interaction (14 -16) manifested in three principal ways: membrane binding, membrane perturbation without permeabilization, and membrane permeabilization (15). Examples of SAPLIP functions include roles in exohydrolase degradation of sphingolipids in the...

show abstract

“…The advantage of using microbial and fungal protease inhibitors is that many of them display unique inhibitory profiles and resistance to proteolytic cleavage, as well as high thermal and broad pH range stability, with the latter being very convenient since harsh conditions may be used for immobilization to the matrix as well as for the several cycles of elution steps, usually involving extreme change in pH and/or ionic strength. The broad-range inhibitor of pepsin-like aspartic proteases, pepstatin A, has been used for purification of aspartic proteases from several different sources, including higher fungi (Sabotič et al 2009a), plants (Payie et al 2000) and insect recombinant enzyme expressed in a bacterial expression system (Volkov et al 2004). Several different types of inhibitor have been used for purification of serine proteases, including the synthetic inhibitor benzamidine and plant-and animal-derived protease inhibitors of different families (Polanowski et al 2003).…”

Section: Applications Of Protease Inhibitors In Biotechnology and Resmentioning

confidence: 99%

Microbial and fungal protease inhibitors—current and potential applications

Sabotič

Kos

2012

Appl Microbiol Biotechnol

145

View full text Add to dashboard Cite

Proteolytic enzymes play essential metabolic and regulatory functions in many biological processes and also offer a wide range of biotechnological applications. Because of their essential roles, their proteolytic activity needs to be tightly regulated. Therefore, small molecules and proteins that inhibit proteases can be versatile tools in the fields of medicine, agriculture and biotechnology. In medicine, protease inhibitors can be used as diagnostic or therapeutic agents for viral, bacterial, fungal and parasitic diseases as well as for treating cancer and immunological, neurodegenerative and cardiovascular diseases. They can be involved in crop protection against plant pathogens and herbivorous pests as well as against abiotic stress such as drought. Furthermore, protease inhibitors are indispensable in protein purification procedures to prevent undesired proteolysis during heterologous expression or protein extraction. They are also valuable tools for simple and effective purification of proteases, using affinity chromatography. Because there are such a large number and diversity of proteases in prokaryotes, yeasts, filamentous fungi and mushrooms, we can expect them to be a rich source of protease inhibitors as well.

show abstract

Untitled

Cited by 4 publications

References 13 publications

Structure and Mechanism of the Pepsin-Like Family of Aspartic Peptidases

Structure and Mechanism of the Pepsin-Like Family of Aspartic Peptidases

Structure and Mechanism of the Saposin-like Domain of a Plant Aspartic Protease

Microbial and fungal protease inhibitors—current and potential applications

Contact Info

Product

Resources

About