Localization of the High‐Affinity ATP Site in Adenosine‐3′:5′‐monophosphate‐Dependent Protein Kinase Type I

Hoppe, Jürgen; Freist, Wolfgang

doi:10.1111/j.1432-1033.1979.tb12804.x

Cited by 28 publications

(3 citation statements)

References 35 publications

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“…In a second step a conformational change takes place in which the adenine binding site is formed. This model is consistent with the assumption of ternary complexes, such as RCcAMP, prior to dissociation of the subunit as proposed and by several groups [28,36,39,46].…”

Section: Discussionsupporting

confidence: 91%

A Model for the Chemical Interactions of Adenosine 3':5'-Monophosphate with the R Subunit of Protein Kinase Type I. Refinement of the Cyclic Phosphate Binding moiety of Protein Kinase Type I

1979

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The cAMP receptor site in the regulatory subunit of adenosine 3' : 5'-monophosphate (CAMP)-dependent protein kinase type I was mapped using analogues of CAMP in which the ribose phosphate moiety was systematically modified. Electronical alteration of the cyclophosphate ring at the 3' and 5' positions by sulfur and nitrogen decreased the affinity of these analogues towards the kinase. Substituents at these positions are not tolerated.Testing the separated diastereomers of derivatives in which one of the exocyclic oxygens at the phosphorus has been substituted by sulfur, it was found that one diastereoisomer is preferentially recognized.Based on these results it is proposed that the hydrophylic cyclic phosphate-ribose moiety of cAMP is bound to the kinase via its 3' and 5'-oxygens, the 2'-hydroxy group and the negative charge in a fixed position.Based on our and other published results it is further proposed, that the adenine moiety is bound in a hydrophobic cleft without any hydrogen bond interactions.The chemical interactions between CAMP and the R subunit of protein kinase type I differ from those found for the binding of cAMP to the chemoreceptor of Dictyostelium discoideum [18].

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Section: Discussionsupporting

confidence: 91%

A Model for the Chemical Interactions of Adenosine 3':5'-Monophosphate with the R Subunit of Protein Kinase Type I. Refinement of the Cyclic Phosphate Binding moiety of Protein Kinase Type I

1979

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“…The question of the origin of the middle T-associated protein kinase activity was approached directly, using two kinds of ATP affinity reagents. 8-Azido-ATP has been used to photolabel the catalytic subunit of protein kinases (25) and ATPases (24). oATP has been used to bind to ATP-utilizing enzymes through the reduction of Schiff bases with borohydride (13,61).…”

Section: The [35s]methionine-labeled Materials Is Largelymentioning

confidence: 99%

Polyoma virus middle T antigen: relationship to cell membranes and apparent lack of ATP-binding activity.

Schaffhausen¹,

Dorai²,

Arakere³

et al. 1982

Mol. Cell. Biol.

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Middle T antigen of polyoma virus is associated principally with the plasma rmembrane. Comparison of the trvpsin sensitivity of middle T in intact cells and "inside out" membrane preparations showed that middle T is oriented towards the inside of the cell. This was confirmed by labeling of middle T in permeabilized cells, but not in intact cells, using [y-32P]ATP. Middle T molecules active in the in vitro kinase reaction could be differentiated from the bulk (metabolically labeled) middle T based on resistance to trypsin treatment. The active fraction also behaved differently from the bulk when cell frameworks were prepared with Triton-containing buffers; whereas the bulk middle T was evenly distributed in the soluble and cell framework fractions, the kinase-active forms were largely associated with the framework. Middle T molecules labeled in vivo with 32P04were found largely in the framework fraction, like the molecules that show kinase activity in vitro. Experiments with ATP affinity reagents 8-azido-ATP and 2,3-dialdehyde ATP have failed to label the middle T antigen. However, 2,3-dialdehyde ATP could be used to inhibit the kinase reaction. This raises the question of whether middle T antigen possesses intrinsic kinase activity or, rather, associates with a cellular tyrosine kinase.

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“…The photoreactive azido analogues of nucleotides have been used to identify the active site of ATPases such as the Na+K +-ATPase of the erythrocyte membrane (Haley and Hoffman, 1974), the F1-ATPase of mitochondria (Wagenvoord et al, 1977;Scheurich et al, 1978), the Ca++-ATPase of the sarcoplasmic reticulum (Briggs et al, 1980), and the 12 S and 18 S dyneins of Chlamydomonas flagella (Ptister et al, 1984;1985). Furthermore, 8-N3ATP has been used to identify the ATP-binding polypeptides of type I protein kinase (Hoppe and Freist, 1979) and the polypeptide associated with the ATPase activity in bovine brain microtubule preparations (Murphy et al, 1983). The preceding experiments have all used purified or partially purified enzymes as substrates in a photoaffinity labeling assay.…”

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confidence: 99%

Identification of a MAP 2-like ATP-binding protein associated with axoplasmic vesicles that translocate on isolated microtubules.

Gilbert¹,

Sloboda²

1986

The Journal of Cell Biology

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Abstract. Axoplasmic vesicles were purified and observed to translocate on isolated microtubules in an ATP-dependent, trypsin-sensitive manner, implying that ATP-binding polypeptides essential for force generation were present on the vesicle surface. To identify these proteins [a32p]8-azidoadenosine 5'-triphosphate ([ct32p]8-N3ATP), a photoaflinity analogue of ATE was used. The results presented here identify and characterize a vesicle-associated polypeptide having a relative molecular mass of 292 kD that bound [a32p]8-N3ATP. The incorporation of label is ultraviolet light-dependent and ATP-sensitive. Moreover, the 292-kD polypeptide could be isolated in association with vesicles or microtubules, depending on the conditions used, and the data indicate that the 292-kD polypeptide is similar to mammalian brain microtubuleassociated protein 2 (MAP 2) for the following reasons: The 292-kD polypeptide isolated from either squid axoplasm or optic lobe cross-reacts with antiserum to porcine brain MAP 2. Furthermore, it purifies with taxol-stabilized microtubules and is released with salt. Based on these characteristics, the 292-kD polypeptide is distinct from the known force-generating molecules myosin and flagellar dynein, as well as the ll0-B0-kD kinesin-like polypeptides that have recently been described (Brady, S. T., 1985, Nature (Lond.), . Because the 292-kD polypeptide binds ATP and is associated with vesicles that translocate on purified MAP-free microtubules in an ATP-dependent fashion, it is therefore believed to be involved in vesicle-microtubule interactions that promote organdie motility. DRECTED intracellular organelle movement is a fundamental process characteristic of all cells. This process has been well studied in the neuron because membranous organelles must travel very long distances, up to a meter or more in length, between the cell body and the synaptic terminal. This process, called fast axonal transport, is ATP-dependent, rapid (1-5 lxm/s), and selective. Newly synthesized membranous organelles produced in the cell body are transported in the orthograde direction toward the nerve terminal. The organelles that are moving in the retrograde direction, however, are different in nature and are being transported from the synaptic terminal region back to the cell body for degradation (reviewed in Grafstein and Forman, 1980;Weiss, 1982). The giant axon of the squid Loligo pealei has been an important preparation for the study of the molecular basis of such organelle transport. The axon is unmyelinated, up to 1 mm in diameter, and 4-8 lxl of axoplasm can be mechanically extruded away from the surrounding plasma membrane and glial sheath (Bear et al., 1937;Lasek, 1974). Moreover, the preparation remains metabolically active for hours, accessible to surrounding solutions for experimentation, and the organelle motility can be observed directly using video-enhanced differential interference contrast microscopy . These characteristics of the axoplasm from Loligo pealei have enhanced the analysis of the relati...

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Localization of the High‐Affinity ATP Site in Adenosine‐3′:5′‐monophosphate‐Dependent Protein Kinase Type I

Cited by 28 publications

References 35 publications

A Model for the Chemical Interactions of Adenosine 3':5'-Monophosphate with the R Subunit of Protein Kinase Type I. Refinement of the Cyclic Phosphate Binding moiety of Protein Kinase Type I

A Model for the Chemical Interactions of Adenosine 3':5'-Monophosphate with the R Subunit of Protein Kinase Type I. Refinement of the Cyclic Phosphate Binding moiety of Protein Kinase Type I

Polyoma virus middle T antigen: relationship to cell membranes and apparent lack of ATP-binding activity.

Identification of a MAP 2-like ATP-binding protein associated with axoplasmic vesicles that translocate on isolated microtubules.

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