Epidermal growth factor dependent phosphorylation of a 35-kilodalton protein in placental membranes

Sheets, Ellen E.; Giugni, Terrence D.; Coates, G. Glenn; Schlaepfer, David D.; Haigler, Harry T.

doi:10.1021/bi00378a026

Cited by 52 publications

(26 citation statements)

References 57 publications

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“…The effect on vesicle aggregation of EGF receptor-catalyzed tyrosine phosphorylation at Tyr-21, on the other hand, has not been elucidated, although it seems clear that this phosphorylation decreases the Ca21 concentration required for phospholipid binding Ando et al, 1989). Moreover, phosphorylation by the EGF receptor converts a certain form of annexin I, which remains associated with membranes_prepared from human placenta in the absence of Ca , into a Ca2+-regulated form whose membrane association is sensitive to chelation of the divalent cation (Sheets et al, 1987). The MVBs of NIH 3T3 cells expressing the human EGF receptor also contain a Ca2 -insensitive form of annexin I, which is rendered Ca2+-sensitive upon phosphorylation by the receptor kinase.…”

Section: Discussionmentioning

confidence: 99%

The association of annexin I with early endosomes is regulated by Ca2+ and requires an intact N-terminal domain.

Seemann

Weber

Osborn

et al. 1996

MBoC

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Annexin I is a member of a multigene family of Ca2+ /phospholipid-binding proteins and a major substrate for the epidermal growth factor (EGF) receptor kinase, which has been implicated in membrane-related events along the endocytotic pathway, in particular in the sorting of internalized EGF receptors occurring in the multivesicular body. We analyzed in detail the intracellular distribution of this annexin by cell fractionation and immunoelectron microscopy. These studies used polyclonal as well as a set of speciesspecific monoclonal antibodies, whose epitopes were mapped to the lateral surface of the molecule next to a region thought to be involved in vesicle aggregation. Unexpectedly, the majority of annexin I was identified on earl and not on multivesicular endosomes in a form that required micromolar levels of Ca + for the association. The specific cofractionation with early endosomes was also observed in transfected baby hamster kidney cells when the intracellular fate of ectopically expressed porcine annexin I was analyzed by using the species-specific monoclonal antibodies in Western blots of subcellular fractions. Interestingly, a truncation of the N-terminal 26, but not the N-terminal 13 residues of annexin I altered its intracellular distribution, shifting it from fractions containing early to those containing late and multivesicular endosomes. These findings underscore the regulatory importance of the N-terminal domain and provide evidence for an involvement of annexin I in early endocytotic processes.

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Section: Discussionmentioning

confidence: 99%

The association of annexin I with early endosomes is regulated by Ca2+ and requires an intact N-terminal domain.

Seemann

Weber

Osborn

et al. 1996

MBoC

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“…This pool of lipocortin is significant as Carnuccio et al (1981) and Cirino & Flower (1987b) reported that it is the external membrane-associated lipocortin 1 that produces its biological effects. Furthermore, the models in which lipocortin 1 has been shown to have biological activity are those in which the protein has been applied externally (Cirino & Flower, 1987a;Cirino et al, 1987;Davidson et al, 1991 (Sheets et al, 1987) and in porcine heart (Pula et al, 1990) and A431 cells (Ando et al, 1991). In the first two examples, lipocortin 1, and 5 and 6 respectively could be extracted by non-ionic detergents but not by EGTA, and hence resemble the membrane-bound pools observed in elicited cells.…”

Section: Discussionmentioning

confidence: 99%

Glucocorticoid — and non‐glucocorticoid induction of lipocortins (annexins) 1 and 2 in rat peritoneal leucocytes in vivo

Peers

Smillie

Elderfield

et al. 1993

British J Pharmacology

107

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1 We have studied the occurrence, distribution and disposition of lipocortins (annexins) 1, 2 and 5 in mixed peritoneal leucocytes obtained from rats in which glucocorticoid levels were altered by adrenalectomy, administration of the glucocorticoid antagonist, RU486, or by injection of dexamethasone or hydrocortisone, as well as from rats in which the peritoneal cells were elicited by inflammatory stimuli. 2 In cells obtained from untreated rats with an intact adrenal cortex, lipocortins 1, 2 and 5 were readily detectable: the majority of each of the proteins was apparently located intracellularly with much smaller amounts in the membrane. Lipocortin 1 and to a lesser extent lipocortin 5 were also seen in a Ca2'-dependent association with the external plasma membrane. Following administration of RU486(2 x 20 mg kg-1) the amounts of lipocortin 1 and 2 in cells were greatly reduced. Conversely, injection of hydrocortisone (1 mg kg-') or dexamethasone (0.08 mg kg-1) caused an increase in the amount of lipocortin 1 and 2 in peritoneal cells within 30 min. Lipocortin 5 was unchanged by any manipulation of glucocorticoid levels. 3 Lipocortins 1 and 2 were elevated in both intracellular and membrane-associated fractions of macrophages elicited by intraperitoneal injection in inflammogens. This phenomenon also occurred in adrenalectomized animals. 4 Our data indicate that glucocorticoids control the synthesis of some members of the lipocortin family in rat mixed peritoneal cells but also suggest the existence of a separate system for controlling the generation of this protein. The significance of these observations is considered in relation to the mechanism of glucocorticoid hormone action on eicosanoid production.

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“…In this report we add placental endonexin II to this growing list. Lipocortin 1 (5,14,15,30) and calpactin I (5) have been shown to be abundant placental proteins and immunological data suggest that endonexin I is also present (D.D.S. and H.T.H., unpublished results).…”

Section: Kpsrlydayelkhalkgagtnekvlteiiasrtpeelraikqvyeewmentioning

confidence: 93%

Structural and functional characterization of endonexin II, a calcium- and phospholipid-binding protein.

Schlaepfer¹,

Mehlman²,

Burgess³

et al. 1987

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

118

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A protein with an apparent Mr of 33,000 was previously purified from the EGTA eluate of a human placental particulate fraction. We now report the amino acid sequence of approximately one-third of this protein and show that it has extensive homology with a newly defined family of Ca2+-binding proteins termed annexins. The partial sequence of the placental protein could be aligned with the sequence of either lipocortin I or calpactin I such that 49% and 58%, respectively, of the residues were identical. A comparison of the partial sequences of the placental protein with the partial sequence of bovine endonexin revealed 74% sequence identity. Based on this close relationship, the placental protein was named endonexin II. Equilibrium dialysis showed that endonexin II bound Ca2+ (Kd >0.5 mM) and the affinity was increased by phosphatidylserine liposomes (Kd 1 i00 jtM Members of this family include proteins initially investigated as substrates for protein-tyrosine kinases, mediators of exocytosis, or components of the cytoskeleton, but their exact physiological roles are not yet known. The realization that they were related was the unexpected result of structural studies. The complete amino acid sequences of two of these proteins, lipocortin I (2) and calpactin I (3-5), have been deduced from cDNA clones: lipocortin I and calpactin I also have been called calpactin 11 (6) and lipocortin II (5), respectively. These proteins have "50% overall sequence homology but have only limited homology (14%) in the N-terminal domain containing 54 amino acids. The Nterminal domains are not required for Ca2+ or phospholipid binding (7,8). The larger core domains contain a 4-fold internal repeat containing a highly conserved 17-amino acid sequence termed the consensus sequence (3, 5). In addition, a bovine protein called endonexin has been partially sequenced and shown to contain four of these consensus regions (9). Partial sequence information of Torpedo calelectrin and protein II shows that these proteins also contain this consensus sequence (9). Other proteins, including calcimedins (10), chromobindins (11), and a variety of proteins with an apparent Mr of ='70,000 (10,11), may be related structurally; however, sequence information is not yet available. None ofthese proteins appears to be related structurally to the Ca2"-binding protein calmodulin or the Ca2+/phospholipid-dependent enzyme protein kinase C.Lipocortin 1 (12-16) and calpactin I (17, 18) are substrates for the protein-tyrosine kinase activities associated with the epidermal growth factor receptor and pp6osrc, respectively, which raises the possibility that these proteins are involved in the regulation of cellular proliferation. However, neither the physiological roles of these proteins nor the effect of phosphorylation on their activities is known. Lipocortin I inhibits phospholipase A2 in an in vitro assay and has been proposed to mediate the steroid-induced antiinflammatory response (2, 19). However, two independent studies have shown that the phospholipase A...

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Epidermal growth factor dependent phosphorylation of a 35-kilodalton protein in placental membranes

Cited by 52 publications

References 57 publications

The association of annexin I with early endosomes is regulated by Ca2+ and requires an intact N-terminal domain.

The association of annexin I with early endosomes is regulated by Ca2+ and requires an intact N-terminal domain.

Glucocorticoid — and non‐glucocorticoid induction of lipocortins (annexins) 1 and 2 in rat peritoneal leucocytes in vivo

Structural and functional characterization of endonexin II, a calcium- and phospholipid-binding protein.

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