Protein Kinases in Copper Homeostasis: A Review on Cu⁺-ATPase Modulation

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

As the author described in the conclusion section, "It is crucial for elucidating the mechanisms underlying various diseases associated with Cu dysregulation, not only in Wilson and Menkes diseases but also in metabolic and neurological disorders, such as Diabetes, Parkinson's, and Alzheimer's." We believe that the most important thing of knowing better the function of kinase or pathway is for serving to understand the pathology and ultimately discover the cure therapies for diseases. So, I suggest please add more detailed content on Cu-associated diseases, such as the mechanisms, the current cure methods, and how to utilize these protein kinases in copper homeostasis to help to find novel therapies. This will raise the significance of the manuscript.

Author Response

Thank you very much for taking the time to review this manuscript. Please find the detailed responses below and the corresponding revisions/corrections highlighted in red in the re-submitted files.

Point-by-point response to Comments and Suggestions for Authors 

Comments 1: As the author described in the conclusion section, "It is crucial for elucidating the mechanisms underlying various diseases associated with Cu dysregulation, not only in Wilson and Menkes diseases but also in metabolic and neurological disorders, such as Diabetes, Parkinson's, and Alzheimer's." We believe that the most important thing of knowing better the function of kinase or pathway is for serving to understand the pathology and ultimately discover the cure therapies for diseases. So, I suggest please add more detailed content on Cu-associated diseases, such as the mechanisms, the current cure methods, and how to utilize these protein kinases in copper homeostasis to help to find novel therapies. This will raise the significance of the manuscript.

Response 1: Thank you for pointing this out. We agree with this suggestion. Therefore, we have inserted information regarding Cu-related diseases in different sections of the manuscript. In “section 1.3.” we added a sentence introducing other pathological conditions associated with Cu homeostasis, such as diabetes, Parkinson’s, Alzheimer’s, and cancer. A complete and extensive review of The FASEB Journal was included (Maung et al., 2021 [29]). (Page number 4, first paragraph, lines 115-118).

In the “Discussion” section, an introductory sentence was inserted in the first paragraph of the discussion (page number 10, second paragraph of the page, lines 429-435) and more detailed information was given as three new paragraphs in the middle of the discussion to emphasize this point (page number 11, second, third and fourth paragraph of page 11, lines 472-496).

At the end of the discussion, a sentence was included to indicate the pharmacological substances used to target cuproptosis (page 11, last paragraph, lines 505-506).

Twelve new references were included to support all new information, with its respective citation numbers:

29. Maung, M. T., Carlson, A., Olea-Flores, M., Elkhadragy, L., Schachtschneider, K. M., Navarro-Tito, N., & Padilla-Benavides, T. The molecular and cellular basis of copper dysregulation and its relationship with human pathologies. FASEB 2021, 35, e21810. https://doi.org/10.1096/fj.202100273RR

63. Gembillo, G.; Labbozzetta, V.; Giuffrida, A.E.; Peritore, L.; Calabrese, V.; Spinella, C.; Stancanelli, M.R.; Spallino, E.; Visconti, L.; Santoro, D. Potential Role of Copper in Diabetes and Diabetic Kidney Disease. Metabolites, 2023, 13, 17. https://doi.org/10.3390/metabo13010017

71. Brady, D. C., Crowe, M. S., Turski, M. L., Hobbs, G. A., Yao, X., Chaikuad, A., Knapp, S., Xiao, K., Campbell, S. L., Thiele, D. J., & Counter, C. M. Copper is required for oncogenic BRAF signalling and tumorigenesis. Nature, 2014, 509, 492–496. https://doi.org/10.1038/nature13180

72. Brady, D. C., Crowe, M. S., Greenberg, D. N., & Counter, C. M. Copper Chelation Inhibits BRAFV600E-Driven Melanomagenesis and Counters Resistance to BRAFV600E and MEK1/2 Inhibitors. Cancer Res., 2017, 77, 6240–6252. https://doi.org/10.1158/0008-5472.CAN-16-1190

73. Xu, M., Casio, M., Range, D. E., Sosa, J. A., & Counter, C. M. Copper Chelation as Targeted Therapy in a Mouse Model of Oncogenic BRAF-Driven Papillary Thyroid Cancer. Clin. Cancer Res., 2018, 24, 4271–4281. https://doi.org/10.1158/1078-0432.CCR-17-3705

75. Wu, K., Chen, L., Kong, Y., Zhuo, J. F., Sun, Q., & Chang, J. The association between serum copper concentration and prevalence of diabetes among US adults with hypertension (NHANES 2011-2016). J. Cell Mol. Med., 2024, 28(8), e18270. https://doi.org/10.1111/jcmm.18270

76. Zhang, S.; Liu, H.; Amarsingh, G.V.; Cheung, C.C.H.; Hogl, S.; Narayanan, U.; Zhang, L.; McHarg, S.; Xu, J.; Gong, D.; et al. Diabetic cardiomyopathy is associated with defective myocellular copper regulation and both defects are rectified by divalent copper chelation. Cardiovasc. Diabetol. 2014, 13, 100. https://doi.org/10.1186/1475-2840-13-100

77. Gong, D., Lu, J., Chen, X., Reddy, S., Crossman, D. J., Glyn-Jones, S., Choong, Y. S., Kennedy, J., Barry, B., Zhang, S., Chan, Y. K., Ruggiero, K., Phillips, A. R., & Cooper, G. J.. A copper(II)-selective chelator ameliorates diabetes-evoked renal fibrosis and albuminuria, and suppresses pathogenic TGF-beta activation in the kidneys of rats used as a model of diabetes. Diabetologia, 2008, 51, 1741–1751. https://doi.org/10.1007/s00125-008-1088-7

78. Farrant, J., Dodd, S., Vaughan, C., Reid, A., Schmitt, M., Garratt, C., Akhtar, M., Mahmod, M., Neubauer, S., Cooper, R. M., Prasad, S. K., Singh, A., Valkovič, L., Raman, B., Ashkir, Z., Clayton, D., Baroja, O., Duran, B., Spowart, C., Bedson, E., … TEMPEST investigators. Rationale and design of a randomised trial of trientine in patients with hypertrophic cardiomyopathy. Heart, 2023, 109, 1175–1182. https://doi.org/10.1136/heartjnl-2022-322271

79. Januzzi, J., Butler, J., Cleland, J., Felker, M., Mentz, R., Wang, Y., Zhang, Y., Mou, H., Yu, J., Guo, L., Li, G., Chen, X., Zhang, J. Effect Of Trientine-hydrochloride In Heart Failure With Lower Left Ventricular Ejection Fraction: The TRACER-HF Trial, J. Card. Fail., 2024, 30, 314–315, https://doi.org/10.1016/j.cardfail.2023.10.472

80. Scholefield, M., Church, S. J., Xu, J., Patassini, S., Roncaroli, F., Hooper, N. M., Unwin, R. D., & Cooper, G. J. S. Widespread Decreases in Cerebral Copper Are Common to Parkinson's Disease Dementia and Alzheimer's Disease Dementia. Front. Aging Neurosci., 2021, 13, 641222. https://doi.org/10.3389/fnagi.2021.641222

86. Pan, Z.; Huang, L.; Gan, Y.; Xia, Y.; Yu,W. The Molecular Mechanisms of Cuproptosis and Small-Molecule Drug Design in Diabetes Mellitus. Molecules, 2024, 29, 2852. https://doi.org/10.3390/molecules29122852

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

The manuscript kinasesphosphatases-3062295  is nice review about the  effects of protein kinases on Cu(I)-ATPases.

Abstract - the construction of the abstract is quite unusual. In my opinion, the aim of the study should be after writing what has been done so far by other authors.

Because the Discussion is quite long, a Conclusion section could be useful.

Please check and rephrase the sentence with high percentage of similarities. 

Minor comments: Table 1 - it would be easier  for the lecturer if the references would be included in the tables for each enzyme. 

Author Response

Thank you very much for taking the time to review this manuscript. Please find the detailed responses below and the corresponding revisions/corrections highlighted in red in the re-submitted files.

Point-by-point response to Comments and Suggestions for Authors

Comments 1: Abstract - the construction of the abstract is quite unusual. In my opinion, the aim of the study should be after writing what has been done so far by other authors.

Response 1: We thank the Reviewer’s suggestion. We have modified the abstract accordingly (page number 1 and lines 10-26).

 

Comments 2:

Because the Discussion is quite long, a Conclusion section could be useful.

Response 2: We appreciate this suggestion. A concise conclusion was provided in the revised manuscript (page number 12  and lines 510-522).

 

Comments 3:

Please check and rephrase the sentence with high percentage of similarities. 

Response 3: We are afraid that we did not completely understand this comment. Is it related to a specific sentence or are there different sentences with a high percentage of similarities? To address this comment, we have checked the manuscript and modified a sentence, as described below:

Original sentence: “This suggests that PKA plays a role in a unique, Cu-dependent trafficking of ATP7A, which differs from what was observed for ATP7B [28]. The Cu-dependent trafficking of ATP7A to the plasma membrane of HeLa cells does not respond to Rho GTPases and PKD, known regulators of carrier vesicles in the TGN [29].”

Revised sentence: “PKA uniquely influences Cu-dependent ATP7A trafficking, unlike ATP7B [28]. In HeLa cells, ATP7A’s Cu-dependent trafficking to the plasma membrane is unaffected by Rho GTPases and PKD, known regulators of carrier vesicles in the TGN [29].” (page 4, fourth paragraph, lines 140-142).

 

Comments 4: Minor comments: Table 1 - it would be easier  for the lecturer if the references would be included in the tables for each enzyme.

Response 4: We acknowledge this observation. We have added new references of seminal articles that describe all enzymes known as cuproenzymes. We decided to use the original papers that describe the presence of Cu in the structure of each enzyme. To be more precise and avoid misunderstandings, we removed catalase and glutathione oxidase from the table. While these enzymes are influenced by Cu, they do not have Cu as a cofactor in their native structure. We apologize for any confusion this may have caused.

All new references included in Table 1 (page 2) are listed below with the respective numbers:

5. Holmberg, C. G., & Laurell, C. B. Investigations in serum copper. II. Isolation of the copper-containing protein, and a description of some of its properties. Acta Chem. Scand., 1948, 5, 476–478. https://doi.org/10.3891/acta.chem.scand.02-0550
6. Vander Wende, C., & Wainio, W. W. The state of the copper in cytochrome c oxidase. J. Biol. Chem., 1960, 235, PC11–PC12. https://doi.org/10.1016/S0021-9258(19)67956-1
7. Udenfriend, S., & Cooper, J. R. The enzymatic conversion of phenylalanine to tyrosine. J. Biol. Chem., 1952, 194, 503–511. https://doi.org/10.1016/S0021-9258(18)55802-6
8. Harris, E. D., & Rayton, J. K. The role of copper in the biosynthesis of lysyl oxidase. Biochem. Biophys. Res. Comm., 1976, 71, 1012–1018. https://doi.org/10.1016/0006-291X(76)90714-3
9.   Eipper, B. A., Milgram, S. L., Husten, E. J., Yun, H. Y., & Mains, R. E. Peptidylglycine α-amidating monooxygenase: a multifunctional protein with catalytic, processing, and routing domains. Protein Science, 1993, 2, 489–497. https://doi.org/10.1002/pro.5560020401
10. McCord, J. M., & Fridovich, I. Superoxide dismutase. An enzymic function for erythrocuprein (hemocuprein). J. Biol. Chem., 1969, 244, 6049–6055. https://doi.org/10.1016/S0021-9258(18)63504-5
11. Marklund S. L. Human copper-containing superoxide dismutase of high molecular weight. PNAS, 1982, 79, 7634–7638. https://doi.org/10.1073/pnas.79.24.7634
12. Matoba, Y., Kumagai, T., Yamamoto, A., Yoshitsu, H., & Sugiyama, M. Crystallographic evidence that the dinuclear copper center of tyrosinase is flexible during catalysis.  J. Biol. Chem., 2006, 281, 8981–8990. https://doi.org/10.1074/jbc.M509785200

Author Response File: Author Response.pdf