Expression and Functional Characterization of the Cancer-related Serine Protease, Human Tissue Kallikrein 14

Borgoño, Carla A.; Michael, Iacovos P.; Shaw, Julie; Luo, Liu‐Ying; Ghosh, Manik C.; Soosaipillai, Antoninus; Grass, Linda; Katsaros, Dionyssios; Diamandis, Eleftherios P.

doi:10.1074/jbc.m608348200

Cited by 104 publications

(106 citation statements)

References 80 publications

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“…The inhibition of NE by SLPI and elafin was used as a positive control. Assays were performed using optimal substrates (PFR-AMC for KLK1; VPR-AMC for KLK5, KLK6, and KLK13; QAR-AMC for KLK14; and AAPV-AMC for NE) and buffer conditions (KLK1: 0.1 M Tris-HCl, 0.1 M NaCl, 0.01% Tween-20, pH 8.0; KLK5, KLK13, and KLK14: 0.1 M sodium phosphate, 0.01% Tween 20, pH 8.0; KLK6: 50 mM Tris, 0.1 M NaCl, 0.2% bovine serum albumin, pH 7.3; and neutrophil elastase: 0.2 M Tris-HCl, 0.01% Tween-20, pH 7.5) (45,(51)(52)(53)(54).…”

Section: Methodsmentioning

confidence: 99%

A Potential Role for Multiple Tissue Kallikrein Serine Proteases in Epidermal Desquamation

Borgoño

Michael

Komatsu

et al. 2007

Journal of Biological Chemistry

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Desquamation of the stratum corneum is a serine proteasedependent process. Two members of the human tissue kallikrein (KLK) family of (chymo)tryptic-like serine proteases, KLK5 and KLK7, are implicated in desquamation by digestion of (corneo)desmosomes and inhibition by desquamation-related serine protease inhibitors (SPIs). However, the epidermal localization and specificity of additional KLKs also supports a role for these enzymes in desquamation. This study aims to delineate the probable contribution of KLK1, KLK5, KLK6, KLK13, and KLK14 to desquamation by examining their interactions, in vitro, with: 1) colocalized SPI, lympho-epithelial Kazal-type-related inhibitor (LEKTI, four recombinant fragments containing inhibitory domains 1-6 (rLEKTI(1-6)), domains 6 -8 and partial domain 9 (rLEKTI(6 -9)), domains 9 -12 (rLEKTI(9 -12)), and domains 12-15 (rLEKTI(12-15)), secretory leukocyte protease inhibitor, and elafin and 2) their ability to digest the (corneo)desmosomal cadherin, desmoglein 1. KLK1 was not inhibited by any SPI tested. KLK5, KLK6, KLK13, and KLK14 were potently inhibited by rLEKTI(1-6), rLEKTI(6 -9), and rLEKTI(9 -12) with K i values in the range of 2.3-28.4 nM, 6.1-221 nM, and 2.7-416 nM for each respective fragment. Only KLK5 was inhibited by rLEKTI(12-15) (K i ‫؍‬ 21.8 nM). No KLK was inhibited by secretory leukocyte protease inhibitor or elafin. Apart from KLK13, all KLKs digested the ectodomain of desmoglein 1 within cadherin repeats, Ca 2؉ binding sites, or in the juxtamembrane region. Our study indicates that multiple KLKs may participate in desquamation through cleavage of desmoglein 1 and regulation by LEKTI. These findings may have clinical implications for the treatment of skin disorders in which KLK activity is elevated.As the outermost layer of the skin, the stratum corneum functions as the body's main protective barrier against physical and chemical damage, dehydration, and microbial pathogens. Inter-corneocyte cohesion within the stratum corneum depends primarily on corneodesmosomes, structurally modified desmosomes (1-3). Akin to classical desmosomes, corneodesmosomes maintain tissue integrity and mediate cell adhesion via calcium-dependent interactions between two families of desmosomal cadherins, the desmogleins (DSG1-4) 2 and desmocollins 1-3 (4, 5). The most abundant isoforms in the stratum corneum include DSG1, DSG4, and desmocollin-1 (6, 7). As specialized desmosomes, corneodesmosomes also contain a unique glycoprotein constituent, corneodesmosin (3).During normal stratum corneum desquamation, the most superficial corneocytes are shed from the skin surface. This process requires proteolysis of the corneodesmosomal adhesion molecules DSG1 (8, 9), desmocollin-1 (10), and corneodesmosin (11) likely mediated by both trypsin-like and chymotrypsin-like serine proteases (9, 12). To date, serine protease activity in the stratum corneum has been attributed to human tissue kallikreins (KLK; encoded by KLK genes (EC 3.4.21)), a subgroup of 15 secreted serine proteases with (chymo)trypsi...

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Section: Methodsmentioning

confidence: 99%

A Potential Role for Multiple Tissue Kallikrein Serine Proteases in Epidermal Desquamation

Borgoño

Michael

Komatsu

et al. 2007

Journal of Biological Chemistry

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259

258

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“…Mature KLK1, produced in a baculovirus/insect cell line system, was kindly provided by Dr. M. Blaber (Florida State University). KLK14 and KLK11 were produced in house, as described previously (30). HUK-IgG, an antibody recognizing KLK1, was kindly provided by Prof. J. Chao (Medical University of South Carolina).…”

Section: Methodsmentioning

confidence: 99%

“…KLK14 has recently been characterized kinetically as a trypsin-like KLK, with substrate preference over P1-Arg (30,31). According to a new report, KLK14 is possibly a key protease in the skin, contributing to approximately half of the total trypsinlike proteolytic activity in the SC layer (32).…”

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confidence: 99%

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Human Kallikrein-related Peptidase 14 (KLK14) Is a New Activator Component of the KLK Proteolytic Cascade

Emami¹,

Diamandis²

2008

Journal of Biological Chemistry

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Human kallikrein-related peptidases (KLKs) are a family of 15 serine proteases mainly known for their biomarker utility in various neoplastic and non-neoplastic diseases. Despite significant progress in understanding their clinical application, little is known about the activation mechanism(s) of this important family of enzymes. Emerging evidence indicates that KLKs are activated in a stepwise manner, which is a characteristic of proteolytic cascades. Thus far, KLK cascades have been implicated in semen liquefaction and skin desquamation. Many members of the KLK family have been reported to be active in seminal plasma and/or skin, suggesting their involvement in common proteolytic cascades. KLK14, in particular, is highly active and has recently been proposed as one of the key trypsin-like proteases involved in skin desquamation. This study aims to elucidate a probable cascade-mediated role of KLK14 by 1) examining KLK14-mediated cleavage of a heptapeptide library encompassing activation sites of the 15 KLKs and 2) verifying activation of certain candidate downstream targets of KLK14 (i.e. pro-KLK1, -KLK3, and -KLK11). Heptapeptides encompassing activation motifs of KLK2, -3, -5, and -11 were cleaved with a high (>85%) cleavage efficiency. Activation of these candidates was confirmed using full-length recombinant proteins. Pro-KLK11, -KLK3, and -KLK1 were rapidly activated in a concentration-dependent manner. Pro-KLK3 regulation was bidirectional because activation was followed by inactivation via internal cleavage of active KLK3. We are proposing a putative cascade model, operating through multiple KLKs. Identification of novel members of such proteolytic cascades will aid in further defining mechanisms involved in seminal/skin homeostasis.Proteases are a major group of enzymes participating in a multitude of physiological processes, including coagulation, apoptosis, and immune responses (1). Due to the irreversible nature of proteolytic activation, proteases often remain as inactive zymogens in quiescent conditions. Activation is often triggered by an external stimulus and mediated by highly orchestrated cascades (2). Within a cascade, proteases function coordinately to ensure sequential activation of protease zymogens. The result is a rapid amplification of signal, in response to a minute amount of stimulus (2).Proteolytic cascades are tightly regulated through a series of highly orchestrated feedback loops, internal cleavages, and (auto)degradations. Also, inhibitors play a major role by targeting activated proteases (2). These multiple regulatory points are critical in preventing deleterious effects due to uncontrolled protease activation. Dysregulated protease activation has been implicated in several pathological conditions, such as amyloidogenesis in Alzheimer disease, intravascular coagulation in sepsis, and desquamation in various skin disorders as well as tumor metastasis, invasion, and angiogenesis in cancer (2-5).KLKs 2 belong to a subgroup of secreted serine proteases within the S1 family of clan ...

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“…dk/services/NetOGlyc/) and structural considerations a single potential O-glycosylation site at Thr144 is present in KLK7, which may account for the partial glycosylation. KLK14 possesses an N-I-S sequon at Asn161 that was apparently not glycosylated upon expression in yeast, insect cells, primate COS-7 cells, and human HEK293 cells (Figure 2) (Brattsand et al, 2005;Borgoño et al, 2007;Rajapakse and Takahashi 2007).…”

Section: The Epidermal Klks 5 7 and 14mentioning

confidence: 99%

Sweetened kallikrein-related peptidases (KLKs): glycan trees as potential regulators of activation and activity

Guo

Skala

Magdolen

et al. 2014

Biological Chemistry

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Most kallikrein-related peptidases (KLKs) are N-glycosylated with N-acetylglucosamine 2 -mannose 9 units at Asn-Xaa-Ser/Thr sequons during protein synthesis and translocation into the endoplasmic reticulum. These N-glycans are modified in the Golgi machinery, where additional O-glycosylation at Ser and Thr takes place, before exocytotic release of the KLKs into the extracellular space. Sequons are present in all 15 members of the KLKs and comparative studies for KLKs from natural and recombinant sources elucidated some aspects of glycosylation. Although glycosylation of mammalian KLKs 1, 3, 4, 6, and 8 has been analyzed in great detail, e.g., by crystal structures, the respective function remains largely unclear. In some cases, altered enzymatic activity was observed for KLKs upon glycosylation. Remarkably, for KLK3/PSA, changes in the glycosylation pattern were observed in samples of benign prostatic hyperplasia and prostate cancer with respect to healthy individuals. Potential functions of KLK glycosylation in structural stabilization, protection against degradation, and activity modulation of substrate specificity can be deduced from a comparison with other glycosylated proteins and their regulation. According to the new concept of protein sectors, glycosylation distant from the active site might significantly influence the activity of proteases. Novel pharmacological approaches can exploit engineered glycans in the therapeutical context.

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Expression and Functional Characterization of the Cancer-related Serine Protease, Human Tissue Kallikrein 14

Cited by 104 publications

References 80 publications

A Potential Role for Multiple Tissue Kallikrein Serine Proteases in Epidermal Desquamation

A Potential Role for Multiple Tissue Kallikrein Serine Proteases in Epidermal Desquamation

Human Kallikrein-related Peptidase 14 (KLK14) Is a New Activator Component of the KLK Proteolytic Cascade

Sweetened kallikrein-related peptidases (KLKs): glycan trees as potential regulators of activation and activity

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