The Role of EF-hand Domains and C2 Domain in Regulation of Enzymatic Activity of Phospholipase Cζ

Kouchi, Zen; Shikano, Tomohide; Nakamura, Yoshikazu; Shirakawa, Hideki; Fukami, Kiyoko; Miyazaki, Shunichi

doi:10.1074/jbc.m412123200

Cited by 110 publications

(117 citation statements)

References 40 publications

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“…The EF-hand domains of most of these enzymes have limited or no ability to bind 4 Ca 2+ (Bairoch and Cox, 1990). A key exception to this is PLCζ, which is responsible for Ca 2+ oscillations in fertilized oocytes and exhibits a high sensitivity to Ca 2+ mediated through its EF-hand domain (Kouchi et al, 2005). Also it appears that the EF-hand domain of PLCδ1 is functionally important and regulates binding of the pleckstrin homology (PH) domain to PIP 2 in a Ca 2+ -dependent manner (Yamamoto et al, 1999).…”

Section: Introductionmentioning

confidence: 99%

A Canonical EF‐Loop Directs Ca²⁺‐Sensitivity in Phospholipase C‐η2

Popovics

Kamil

Morgan

et al. 2014

J of Cellular Biochemistry

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Phospholipase C-ɳ(PLCɳ) enzymes are a class of phosphatidylinositol 4,5-bisphosphatehydrolyzing enzymes involved in intracellular signaling. PLCɳ2 can sense Ca 2+ (stimulated bỹ 1 µM free Ca 2+ ) suggesting that it can amplify transient Ca 2+ signals. PLCɳ enzymes possess an EF-hand domain composed of two EF-loops; a canonical 12-residue loop (EF-loop 1) and a non-canonical 13-residue loop (EF-loop 2). Ca 2+ -binding to synthetic peptides corresponding to EF-loops 1 and 2 of PLCɳ2 and EF-loop 1 of calmodulin (as a control) was examined by 2D-[ 1 H, 1 H] TOCSY NMR. Both PLCɳ2 EF-loop peptides bound Ca 2+ in a similar manner to that of the canonical calmodulin EF-loop 1, particularly at their N-terminus. A molecular model of the PLCɳ2 EF-hand domain, constructed based upon the structure of calmodulin, suggested both EF-loops may participate in Ca 2+ -binding. To determine whether the EF-hand is responsible for Ca 2+ -sensing, inositol phosphate accumulation was measured in COS7 cells transiently expressing wild-type or mutant PLCɳ2 proteins. Addition of 70µM monensin (a Na + /H + antiporter that increases intracellular Ca 2+ ) induced a 4 to 7-fold increase in wild-type PLCɳ2 activity. In permeabilized cells, PLCn2 exhibited a ~4-fold increase in activity in the presence of 1 µM free Ca 2+ . The D256A (EF-loop1) mutant exhibited a ~10-fold reduction in Ca 2+ -sensitivity and was not activated by monensin, highlighting the involvement of EF-loop 1 in Ca 2+ -sensing. Involvement of EF-loop 2 was examined using D292A, H296A, Q297A and E304A mutants. Interestingly, the monensin responses and Ca 2+ -sensitivities were largely unaffected by the mutations, indicating that the non-canonical EF-loop 2 is not involved in Ca 2+ -sensing.

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Section: Introductionmentioning

confidence: 99%

A Canonical EF‐Loop Directs Ca²⁺‐Sensitivity in Phospholipase C‐η2

Popovics

Kamil

Morgan

et al. 2014

J of Cellular Biochemistry

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“…A study investigating the function of the domains in mouse Plcz1 has confirmed that the N-terminal EF-hand domains, especially EF-1 and EF-3, are essential for the function of Plcz1. The lack of the three EF-hand domains has been shown to result in a low level of Ca 2+ sensitivity [17] and loss of the Ca 2+ oscillations associated with fertilization in mouse eggs injected with EF domain RNA. This is in contrast to the activity of the full-length Plcz1, which can trigger Ca 2+ oscillations in mouse eggs [18][19][20].…”

Section: Discussionmentioning

confidence: 99%

NYD-SP27, a novel intrinsic decapacitation factor in sperm

Bi¹,

Xu²,

Wong³

et al. 2009

Asian J Androl

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Abstract:Prior to fertilization sperm has to undergo an activation process known as capaciation, leading to the acrosome reaction. Till now, little is known about the mechanism for preventing premature capacitation in sperm although decapacitation factors from various sources have been thought to be involved. In this study, we report that NYD-SP27, an isoform of phospholipase C Zeta 1 (PLCZ1), is localized to the sperm acrosome in mouse and human spermatozoa by immunofluorescence using a specific antibody. Western blot and double staining analyses show NYD-SP27 becomes detached from sperm, as they undergo capacitation and acrosome reaction. The absence of HCO 3 -, a key factor in activating capacitation, from the capacitation-inducing medium prevents the loss of NYD-SP27 from sperm. The anti-NYD-SP27 antibody also prevents the loss of NYD-SP27 from sperm, reduced the number of capacitated sperm, inhibited the acrosome reaction induced by ATP and progesterone, and inhibited agonist-induced PLC-coupled Ca 2+ mobilization in sperm, which can be mimicked by the PLC inhibitor, U73122. These data strongly suggest that NYD-SP27 is a physiological inhibitor of PLC that acts as an intrinsic decapacitation factor in sperm to prevent premature capacitation and acrosome reaction.

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“…La PLC a la particularité d'être extrêmement sensible au calcium, et on pense qu'elle est naturellement active aux concentrations cytosoliques de calcium (10-100 nM) [36]. Cependant, dans le spermatozoïde, elle serait inactivée par la fixation de son domaine C2 (calcium-binding motif) à certains phosphatidylinositides comme le PIP3 et le PIP5 [37].…”

Section: Rôles Des Phospholipases Dans La Réaction Acrosomiqueunclassified

Les phospholipases, enzymes clés de la physiologie spermatique

et al. 2012

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> Chez les mammifères, la physiologie du spermatozoïde dans les voies génitales femelles est complexe, et plusieurs étapes bien définies doivent être franchies pour que le spermatozoïde puisse féconder l'ovocyte. Les phospholipases, qui permettent de contrôler la composition lipidique du spermatozoïde en hydrolysant les phospholipides de manière très spécifique, sont impliquées dans des mécanismes aussi variés que le contrôle du battement flagellaire, la capacitation, la réaction acrosomique, ainsi que dans le déclenchement des oscillations calciques de l'ovocyte qui entraînent l'activation ovocytaire. Deux phospholipases, la PLC et la PLA 2 sécrétée de groupe X, pourraient représenter des cibles thérapeutiques pour lutter contre l'infertilité masculine. < constituées d'une tête polaire phosphoalcoolique reliée, via une molécule de glycérol, à une queue hydrophobe constituée de deux acides gras [1]. Choline et éthanolamine sont les rési-dus alcool les plus courants au niveau de la partie polaire, mais d'autres, tels que la sérine ou l'inositol biphosphate, composent certaines GPL (Figure 1). Tous ces GPL sont hydrolysés par une superfamille de protéines, les phospholipases (PL). Les PL sont sépa-rées en quatre familles sans homologie structurale, sur la base de la localisation du site enzymatique d'hydrolyse :

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The Role of EF-hand Domains and C2 Domain in Regulation of Enzymatic Activity of Phospholipase Cζ

Cited by 110 publications

References 40 publications

A Canonical EF‐Loop Directs Ca²⁺‐Sensitivity in Phospholipase C‐η2

A Canonical EF‐Loop Directs Ca²⁺‐Sensitivity in Phospholipase C‐η2

NYD-SP27, a novel intrinsic decapacitation factor in sperm

Les phospholipases, enzymes clés de la physiologie spermatique

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The Role of EF-hand Domains and C2 Domain in Regulation of Enzymatic Activity of Phospholipase Cζ

Cited by 110 publications

References 40 publications

A Canonical EF‐Loop Directs Ca2+‐Sensitivity in Phospholipase C‐η2

A Canonical EF‐Loop Directs Ca2+‐Sensitivity in Phospholipase C‐η2

NYD-SP27, a novel intrinsic decapacitation factor in sperm

Les phospholipases, enzymes clés de la physiologie spermatique

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Product

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A Canonical EF‐Loop Directs Ca²⁺‐Sensitivity in Phospholipase C‐η2

A Canonical EF‐Loop Directs Ca²⁺‐Sensitivity in Phospholipase C‐η2