Peflin and ALG-2, Members of the Penta-EF-Hand Protein Family, Form a Heterodimer That Dissociates in a Ca2+-dependent Manner

Kitaura, Yasuyuki; Matsumoto, Shinji; Satoh, Hiroyuki; Hitomi, Kiyotaka; Maki, Masatoshi

doi:10.1074/jbc.m008649200

Cited by 79 publications

(113 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Treatment of these cells with thapsigargin, a drug known to release calcium from intracellular stores and cause cell death through ER stress with potential ALG-2 involvement (2), did not result in any change in localization of ALG-2. The contrast in findings between our studies and others (21,28,29) may be due to the different cancer cell types being examined or the fidelity of the antibodies used in the studies (30).Various biochemical and structural approaches have been used to characterize different ALG-2 analogues (3,4,12,18,20,31) . As with other members of this family of Ca 2+ -binding proteins, ALG-2 exists as a dimer in solution (32,33).…”

mentioning

confidence: 56%

See 1 more Smart Citation

Ca²⁺ Binding to EF Hands 1 and 3 Is Essential for the Interaction of Apoptosis-Linked Gene-2 with Alix/AIP1 in Ocular Melanoma

et al. 2004

View full text Add to dashboard Cite

Apoptosis-linked gene-2 (ALG-2) encodes a 22 kDa Ca 2+ -binding protein of the penta EF-hand family that is required for programmed cell death in response to various apoptotic agents. Here, we demonstrate that ALG-2 mRNA and protein are down-regulated in human uveal melanoma cells compared to their progenitor cells, normal melanocytes. The down regulation of ALG-2 may provide melanoma cells with a selective advantage. ALG-2 and its putative target molecule, Alix/AIP1, are localized primarily in the cytoplasm of melanocytes and melanoma cells independent of the intracellular Ca 2+ concentration or the activation of apoptosis. Cross-linking and analytical centrifugation studies support a single-species dimer conformation of ALG-2, also independent of Ca 2+ concentration. However, binding of Ca 2+ to both EF-1 and EF-3 is necessary for ALG-2 interaction with Alix/AIP1 as demonstrated using surface plasmon resonance spectroscopy. Mutations in EF-5 result in reduced target interaction without alteration in Ca 2+ affinity. The addition of N-terminal ALG-2 peptides, residues 1-22 or residues 7-17, does not alter the interaction of ALG-2 or an N-terminal deletion mutant of ALG-2 with Alix/AIP1, as might be expected from a model derived from the crystal structure of ALG-2. Fluorescence studies of ALG-2 demonstrate that an increase in surface hydrophobicity is primarily due to Ca 2+ binding to EF-3, while Ca 2+ binding to EF-1 has little effect on surface exposure of hydrophobic residues. Together, these data indicate that gross surface hydrophobicity changes are insufficient for target recognition. † This work was supported by NIH Grants EY12768 and EY13705 (A.S.P.), EY08061 (K.P.), and EY06603 and EY14239 (J.W.C.) and the Swiss National Science Foundation 31-65071.01 (J.C.), as well as grants from the Retina Research Foundation, Research to Prevent Blindness Inc. (RPB), the University of Wisconsin Comprehensive Cancer Center, and the E. K. Bishop Foundation. L.S. is a recipient of the Cremer Scholarship. A.S.P. is a Jules and Doris Stein RPB Professor. * To whom correspondence should be addressed. NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author ManuscriptApoptosis-linked gene-2 (ALG-2) was first identified in a death-trap assay using a mouse T cell hybridoma model (1). These cells when transfected with an antisense ALG-2 expression vector were resistant to T cell receptor-mediated cell death and were partially protected from other agents that normally initiate programmed cell death such as Fas, staurosporine, actinomycin D, and C 2 -ceramide (1). More recent work suggests that ALG-2 might be involved in the apaf1-independent intrinsic apoptotic pathway activated as a result of endoplasmic reticulum (ER) stress (2).ALG-2 contains five putative EF-hand sequences and is a member of the family of PEF 1 (penta-EF-hand) proteins that includes the calpain small subunit, sorcin, and grancalcin (3). In ALG-2, EF-1 and EF-3 are considered the high-affinity Ca 2+ -binding sites, while one low-affinity site...

show abstract

mentioning

confidence: 56%

“…Various biochemical and structural approaches have been used to characterize different ALG-2 analogues (3,4,12,18,20,31) . As with other members of this family of Ca 2+ -binding proteins, ALG-2 exists as a dimer in solution (32,33).…”

Section: Discussionmentioning

confidence: 99%

Ca²⁺ Binding to EF Hands 1 and 3 Is Essential for the Interaction of Apoptosis-Linked Gene-2 with Alix/AIP1 in Ocular Melanoma

et al. 2004

View full text Add to dashboard Cite

show abstract

“…PDCD6 is required for T cell receptor-, glucocorticoid-(26), and FAS-(28) induced cell death. It interacts with the SH3-binding domain containing pro-apoptotic protein AIP1 (ALG-2-interacting protein-1) (29), peflin (30), and annexin XI (31) in a Ca 2ϩ -dependent way as well as with DAPK1 (deathassociated protein kinase 1) (32). During FAS-induced apoptosis, PDCD6, which is a 22-kDa protein, is cleaved in its N-terminus to yield a 19-kDa protein and translocates from the cytoplasmic membrane to the cytosol (28).…”

Section: Discussionmentioning

confidence: 99%

Genomic markers for malignant progression in pulmonary adenocarcinoma with bronchioloalveolar features

Aviel‐Ronen

Coe

Lau

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Bronchioloalveolar carcinoma (BAC), a subtype of lung adenocarcinoma (ADC) without stromal, vascular, or pleural invasion, is considered an in situ tumor with a 100% survival rate. However, the histological criteria for invasion remain controversial. BAC-like areas may accompany otherwise invasive adenocarcinoma, referred to as mixed type adenocarcinoma with BAC features (AWBF). AWBF are considered to evolve from BAC, representing a paradigm for malignant progression in ADC. However, the supporting molecular evidence remains forthcoming. Here, we have studied the genomic changes of BAC and AWBF by array comparative genomic hybridization (CGH). We used submegabaseresolution tiling set array CGH to compare the genomic profiles of 14 BAC or BAC with focal area suspicious for invasion with those of 15 AWBF. Threshold-filtering and frequency-scoring analysis found that genomic profiles of noninvasive and focally invasive BAC are indistinguishable and show fewer aberrations than tumor cells in BAC-like areas of AWBF. These aberrations occurred mainly at the subtelomeric chromosomal regions. Increased genomic alterations were noted between BAC-like and invasive areas of AWBF. We identified 113 genes that best differentiated BAC from AWBF and were considered candidate marker genes for tumor invasion and progression. Correlative gene expression analyses demonstrated a high percentage of them to be poor prognosis markers in early stage ADC. Quantitative PCR also validated the amplification and overexpression of PDCD6 and TERT on chromosome 5p and the prognostic significance of PDCD6 in early stage ADC patients. We identified candidate genes that may be responsible for and are potential markers for malignant progression in AWBF.array comparative genomic hybridization ͉ bronchioloalveolar carcinoma ͉ microarray ͉ non-small-cell lung carcinoma ͉ prognostic markers

show abstract

“…The x-ray structures show that three EF sites are potentially able to bind Ca 2ϩ in the physiological range of concentrations, namely the unusual EF1 motif and EF2 and EF3, whose structural features resemble those of EF-hands in regulatory Ca 2ϩ -binding proteins. However, binding of only two Ca 2ϩ /monomer suffices to trigger the conformational change that exposes hydrophobic regions and leads to interaction with the respective targets (3,19,20). The available data indicate that functional coupling does not involve the same EF-hands in the different proteins, suggesting that despite the high structural similarity there is no common mode of information transfer within the PEF family.…”

mentioning

confidence: 89%

Information Transfer in the Penta-EF-hand Protein Sorcin Does Not Operate via the Canonical Structural/Functional Pairing

Mella

Colotti

Zamparelli

et al. 2003

Journal of Biological Chemistry

View full text Add to dashboard Cite

Sorcin is a typical penta-EF-hand protein that participates in Ca2؉ -regulated processes by translocating reversibly from cytosol to membranes, where it interacts with different target proteins in different tissues. Binding of two Ca 2؉ /monomer triggers translocation, although EF1, EF2, and EF3 are potentially able to bind calcium at micromolar concentrations. To identify the functional pair, the conserved bidentate -Z glutamate in these EF-hands was mutated to yield E53Q-, E94A-, and E124A-sorcin, respectively. Limited structural perturbations occur only in E124A-sorcin due to involvement of Glu-124 in a network of interactions that comprise the long D helix connecting EF3 to EF2. The overall affinity for Ca 2؉ and for two sorcin targets, annexin VII and the ryanodine receptor, follows the order wildtype > E53Q-> E94A-> E124A-sorcin, indicating that disruption of EF3 has the largest functional impact and that disruption of EF2 and EF1 has progressively smaller effects. Based on this experimental evidence, EF3 and EF2, which are not paired in the canonical manner, are the functional EF-hands. Sorcin is proposed to be activated upon Ca 2؉ binding to EF3 and transmission of the conformational change at Glu-124 via the D helix to EF2 and from there to EF1 via the canonical structural/functional pairing. This mechanism may be applicable to all penta-EF-hand proteins.

show abstract

Peflin and ALG-2, Members of the Penta-EF-Hand Protein Family, Form a Heterodimer That Dissociates in a Ca2+-dependent Manner

Abstract: Peflin, a newly identified 30-kDa Ca 2؉ -binding protein, belongs to the penta-EF-hand (PEF) protein family, which includes the calpain small subunit, sorcin, grancalcin, and ALG-2 (apoptosis-linked gene 2). We pre-

Cited by 79 publications

References 28 publications

Ca²⁺ Binding to EF Hands 1 and 3 Is Essential for the Interaction of Apoptosis-Linked Gene-2 with Alix/AIP1 in Ocular Melanoma

Ca²⁺ Binding to EF Hands 1 and 3 Is Essential for the Interaction of Apoptosis-Linked Gene-2 with Alix/AIP1 in Ocular Melanoma

Genomic markers for malignant progression in pulmonary adenocarcinoma with bronchioloalveolar features

Information Transfer in the Penta-EF-hand Protein Sorcin Does Not Operate via the Canonical Structural/Functional Pairing

Contact Info

Product

Resources

About

Peflin and ALG-2, Members of the Penta-EF-Hand Protein Family, Form a Heterodimer That Dissociates in a Ca2+-dependent Manner

Abstract: Peflin, a newly identified 30-kDa Ca 2؉ -binding protein, belongs to the penta-EF-hand (PEF) protein family, which includes the calpain small subunit, sorcin, grancalcin, and ALG-2 (apoptosis-linked gene 2). We pre-

Cited by 79 publications

References 28 publications

Ca2+ Binding to EF Hands 1 and 3 Is Essential for the Interaction of Apoptosis-Linked Gene-2 with Alix/AIP1 in Ocular Melanoma

Ca2+ Binding to EF Hands 1 and 3 Is Essential for the Interaction of Apoptosis-Linked Gene-2 with Alix/AIP1 in Ocular Melanoma

Genomic markers for malignant progression in pulmonary adenocarcinoma with bronchioloalveolar features

Information Transfer in the Penta-EF-hand Protein Sorcin Does Not Operate via the Canonical Structural/Functional Pairing

Contact Info

Product

Resources

About

Ca²⁺ Binding to EF Hands 1 and 3 Is Essential for the Interaction of Apoptosis-Linked Gene-2 with Alix/AIP1 in Ocular Melanoma

Ca²⁺ Binding to EF Hands 1 and 3 Is Essential for the Interaction of Apoptosis-Linked Gene-2 with Alix/AIP1 in Ocular Melanoma