• Interaction

    AccNo. 22515 Score 0.86
    Name ARASE_hK2 Environment in vitro
    Kd 1.0 Organism N/A
    Function peptide is a substrate of kallikrein 2


    AccNo. 22399
    Name ARASE
    Organism N/A
    Constraint none
    Sequence ARASE
    Origin phage display
    Form phage
    Internalized no
    Unnatural no
    Imaging no
    Is Motif no


    AccNo. 22250
    Name hK2
    Description kallikrein 2
    Organism Homo sapiens (human)
    Type protein


    AccNo. 22468
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    Name PD_6
    Detection filamentous phage display, MI:0048
    Source PDF Text Id 6
    Journal Eur J Biochem. 2002 Jun;269(11):2747-54.
    Title Substrate specificity of human kallikrein 2 (hK2) as determined by phage display technology.
    Authors Cloutier SM, Chagas JR, Mach JP, Gygi CM, Leisinger HJ, Deperthes D
    Text Human glandular kallikrein 2 (hK2) is a trypsin-like serine protease expressed predominantly in the prostate epithelium. Recently, hK2 has proven to be a useful marker that can be used in combination with prostate specific antigen for screening and diagnosis of prostate cancer. The cleavage by hK2 of certain substrates in the proteolytic cascade suggest that the kallikrein may be involved in prostate cancer development; however, there has been very little other progress toward its biochemical characterization or elucidation of its true physiological role. In the present work, we adapt phage substrate technology to study the substrate specificity of hK2. A phage-displayed random pentapeptide library with exhaustive diversity was generated and then screened with purified hK2. Phages displaying peptides susceptible to hK2 cleavage were amplified in eight rounds of selection and genes encoding substrates were transferred from the phage to a fluorescent system using cyan fluorescent protein (derived from green fluorescent protein) that enables rapid determination of specificity constants. This study shows that hK2 has a strict preference for Arg in the P1 position, which is further enhanced by a Ser in P'1 position. The scissile bonds identified by phage display substrate selection correspond to those of the natural biological substrates of hK2, which include protein C inhibitor, semenogelins, and fibronectin. Moreover, three new putative hK2 protein substrates, shown elsewhere to be involved in the biology of the cancer, have been identified thus reinforcing the importance of hK2 in prostate cancer development.
    Mesh Terms Green Fluorescent Proteins; Humans; Kinetics; Luminescent Proteins; Peptide Library; Substrate Specificity/genetics; Tissue Kallikreins/genetics; Tissue Kallikreins/metabolism
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