Copper complexes of glycyl-histidyl-lysine and two of its synthetic analogues: Chemical behaviour and biological activity
Abstract
Copper complex formation equilibria of glycyl-L-histidyl-L-lysine (Gly-His-Lys, GHK) and of two synthetic analogues, where the histidine residue was replaced with a synthetic amino acid (L-spinacine or L-1,2,3,4-tetrahydro-isoquinoline-3-carboxylic acid), have been carefully investigated using different experimental techniques: potentiometry, solution calorimetry, UV-VIS spectrophotometry, circular dichroism and electron paramagnetic resonance spectroscopies. All the ligands formed complexes having different stoichiometries and stabilities; evidence for the formation of binuclear species is also shown. The structures of the main complexes are discussed. It is suggested that the lateral lysine amino group participates in complex formation, but only at alkaline pH values: at physiological pH this group is protonated and available for possible interactions with cellular receptors. The above tripeptides have been tested for their enzymatic stability in human serum: the synthetic compounds showed no significant degradation for at least 3 h. Finally, their activity as growth factor has been studied in vitro. The two synthetic analogues showed an activity comparable to or even higher than that of GHK, thus suggesting their possible use as additives in cell culture media, even in the presence of serum. Relevant information on the GHK action mechanism as cell growth factor has been obtained: the formation of copper complexes, driven by the first (Gly) residue, appears necessary while the second residue (His) does not appear to play a specific role; the presence of the free side chain of the third residue (Lys) appears to be of fundamental importance.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
The paper's calculated protonation and formation constants are closest to those obtained by Lau et al. and Conato et al., two groups that also utilised potentiometry.
“Our calculated protonation and formation constants values are closest to those obtained by Lau et al. [28] and Conato et al. [18], two groups that also utilised potentiometry to calculate the binding constants.”
From Ternary Cu(II) Complex with GHK Peptide and Cis-Urocanic Acid as a Potential Physiologically Functional Copper Chelate (2020) · cites this paper as [18]
The cK7.4 value for Cu(GHK)2 is significantly lower than the value ca. 500 M−1 that could be inferred from potentiometric stability constants reported by Conato et al.
“This value is significantly lower from the value ca. 500 M−1 that could be inferred from potentiometric stability constants reported by Conato et al. [18].”
From Ternary Cu(II) Complex with GHK Peptide and Cis-Urocanic Acid as a Potential Physiologically Functional Copper Chelate (2020) · cites this paper as [18]
Cites
Maquart, F.-X., Pickart, L., Laurent, M., Gillery, P., Monboisse, J.-C., & Borel, J.-P. (1988) · FEBS Letters · doi:10.1016/0014-5793(88)80509-x · Cited by 181 · see citing works
Growth-modulating plasma tripeptide may function by facilitating copper uptake into cells
Pickart, L., Freedman, J. H., Loker, W. J., Peisach, J., Perkins, C. M., Stenkamp, R. E., & Weinstein, B. (1980) · Nature · doi:10.1038/288715a0 · Cited by 175 · see citing works
Lau, S. J., & Sarkar, B. (1981) · Biochemical Journal · doi:10.1042/bj1990649 · Cited by 79 · see citing works
Daniele, P. G., Zerbinati, O., Zelano, V., & Ostacoli, G. (1991) · J. Chem. Soc., Dalton Trans. · doi:10.1039/dt9910002711 · Cited by 49 · see citing works
Flagged first, then by citations
Cited by
Sorted by citations
Chemical and biological aspects of Cu2+ interactions with peptides and aminoglycosides
Kozłowski, H., Kowalik‐Jankowska, T., & Jeżowska‐Bojczuk, M. (2005) · Coordination Chemistry Reviews · doi:10.1016/j.ccr.2005.04.027 · Cited by 169 · see citing works
Self-citationHureau, C., Eury, H., Guillot, R., Bijani, C., Sayen, S., Solari, P., Guillon, E., Faller, P., & Dorlet, P. (2011) · Chemistry · doi:10.1002/chem.201100751 · Cited by 122 · see citing works
Engineering metal ion coordination to regulate amyloid fibril assembly and toxicity
Dong, J., Canfield, J. M., Mehta, A. K., Shokes, J. E., Tian, B., Childers, W. S., Simmons, J. A., Mao, Z., Scott, R. A., Warncke, K., et al. (2007) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0702669104 · Cited by 133 · see citing works
Gonzalez, P., Bossak‐Ahmad, K., Stefaniak, E., Hureau, C., Raibaut, L., Bal, W., & Faller, P. (2018) · Chemistry - A European Journal · doi:10.1002/chem.201705398 · Cited by 142 · see citing works
Trapaidze, A., Hureau, C., Bal, W., Winterhalter, M., & Faller, P. (2012) · JBIC Journal of Biological Inorganic Chemistry · doi:10.1007/s00775-011-0824-5 · Cited by 103 · see citing works
Valenti, L. E., de Pauli, C. D., & Giacomelli, C. (2006) · Journal of Inorganic Biochemistry · doi:10.1016/j.jinorgbio.2005.11.003 · Cited by 73
Copper Chelators: Chemical Properties and Bio-medical Applications
Tegoni, M., Valensin, D., Toso, L., & Remelli, M. (2014) · Current Medicinal Chemistry · doi:10.2174/0929867321666140601161939 · Cited by 56 · see citing works
Self-citationLights and Shadows on the Therapeutic Use of Antimicrobial Peptides
Bellotti, D., & Remelli, M. (2022) · Molecules · doi:10.3390/molecules27144584 · Cited by 50 · see citing works
“In the presence of proteases, the half-life of peptides is generally of the order of minutes, only rarely of a few hours [38,99].”
BackgroundSelf-citationHuman skin penetration of a copper tripeptide in vitro as a function of skin layer
Hostynek, J. J., Dreher, F., & Maibach, H. I. (2011) · Inflammation Research · doi:10.1007/s00011-010-0238-9 · Cited by 47 · see citing works
Jakab, N. I., Gyurcsik, B., Körtvélyesi, T., Vosekalna, I., Jensen, J. H., & Larsen, E. (2007) · Journal of Inorganic Biochemistry · doi:10.1016/j.jinorgbio.2007.05.012 · Cited by 45 · see citing works
Peptides as Active Ingredients: A Challenge for Cosmeceutical Industry
Ledwoń, P., Errante, F., Papini, A., Rovero, P., & Latajka, R. (2020) · Chemistry and Biodiversity · doi:10.1002/cbdv.202000833 · Cited by 40 · see citing works
Mazurowska, L., & Mojski, M. (2009) · Journal of the Society of Cosmetic Chemists · doi:10.1111/j.1468-2494.2008.00446_5.x · Cited by 38 · see citing works
Lee, S., Lee, S. M., Lee, S. H., Choi, W.-K., Park, S. J., Kim, D. Y., Oh, S. W., Oh, J., Cho, J. Y., Lee, J., et al. (2023) · Acta Biomaterialia · doi:10.1016/j.actbio.2023.10.011 · Cited by 38 · see citing works
Lakatos, A., Gyurcsik, B., May, N. V., Csendes, Z., Wéber, E., Fülöp, L., & Kiss, T. (2011) · Dalton Transactions · doi:10.1039/c1dt10989h · Cited by 39 · see citing works
Roviello, G., Musumeci, D., Bucci, E., & Pedone, C. (2011) · Molecular Biosystems · doi:10.1039/c0mb00214c · Cited by 33 · see citing works
Ufnalska, I., Drew, S. C., Zhukov, I., Szutkowski, K., Wawrzyniak, U. E., Wróblewski, W., Frączyk, T., & Bal, W. (2021) · Inorganic Chemistry · doi:10.1021/acs.inorgchem.1c02669 · Cited by 35 · see citing works
Bossak-Ahmad, K., Wiśniewska, M. D., Bal, W., Drew, S. C., & Frączyk, T. (2020) · International Journal of Molecular Sciences · doi:10.3390/ijms21176190 · Cited by 25 · see citing works
“[18], two groups that also utilised potentiometry to calculate the binding constants.”
BackgroundGonzalez, P., Vileno, B., Bossak‐Ahmad, K., Khoury, Y. E., Hellwig, P., Bal, W., Hureau, C., & Faller, P. (2017) · Inorganic Chemistry · doi:10.1021/acs.inorgchem.7b01996 · Cited by 31 · see citing works
Conato, C., Kozłowski, H., Młynarz, P., Pulidori, F., & Remelli, M. (2002) · Polyhedron · doi:10.1016/s0277-5387(02)00952-x · Cited by 31 · see citing works
Self-citationBossak, K., Mital, M., Poznański, J., Bonna, A., Drew, S., & Bal, W. (2016) · Inorganic Chemistry · doi:10.1021/acs.inorgchem.6b01441 · Cited by 23 · see citing works
Chikira, M. (2008) · Journal of Inorganic Biochemistry · doi:10.1016/j.jinorgbio.2008.01.019 · Cited by 23
MAZUROWSKA, L., & MOJSKI, M. (2007) · Talanta · doi:10.1016/j.talanta.2006.11.034 · Cited by 23 · see citing works
Hostynek, J. J., Dreher, F., & Maibach, H. I. (2010) · Inflammation Research · doi:10.1007/s00011-010-0214-4 · Cited by 22 · see citing works
“It was not analyzed for stoichiometric purity since, in several studies, copper complexes in a physiological environment were subject to de-complexation and rearrangement, depending on pH and complex stability constants [15, 16].”
BackgroundHsiao, C., Wu, H.-H., Malhotra, N., Liu, Y.-C., Wu, Y.-H., Lin, Y.-N., Saputra, F., Santoso, F., & Chen, K. H. ‐. (2020) · Biomolecules · doi:10.3390/biom10091202 · Cited by 23 · see citing works
“Thus, the GHK tripeptide, as a modulator for cellular growth, can play a versatile role in the biomedical and biotechnological development of new drugs, with potential applications [12].”
BackgroundCu(ii) ion coordination to the pentadecapeptide model of the SPARC copper-binding site
Conato, C., Kamysz, W., Kozłowski, H., Łuczkowski, M., Maćkiewicz, Z., Młynarz, P., Remelli, M., Valensin, D., & Valensin, G. (2002) · Journal of the Chemical Society Dalton Transactions · doi:10.1039/b207029d · Cited by 26 · see citing works
Self-citationSchirer, A., Khoury, Y. E., Faller, P., & Hellwig, P. (2017) · JBIC Journal of Biological Inorganic Chemistry · doi:10.1007/s00775-017-1449-0 · Cited by 19 · see citing works
Matera-Witkiewicz, A., Brasuń, J., Świątek-Kozłowska, J., Pratesi, A., Ginanneschi, M., & Messori, L. (2009) · Journal of Inorganic Biochemistry · doi:10.1016/j.jinorgbio.2009.01.001 · Cited by 18
The possible role of Gly residues in the prion octarepeat region in the coordination of Cu2+ ions
Luczkowski, M., Kozłowski, H., Łęgowska, A., Rolka, K., & Remelli, M. (2003) · doi:10.1039/b207702g · Cited by 18
Self-citationBadenhorst, T., Svirskis, D. M., & Wu, Z. (2014) · Pharmaceutical Development and Technology · doi:10.3109/10837450.2014.979944 · Cited by 17 · see citing works
Cu(ii) ion coordination to SPARC: a model study on short peptide fragments
Remelli, M., Łuczkowski, M., Bonna, A., Maćkiewicz, Z., Conato, C., & Kozłowski, H. (2003) · New Journal of Chemistry · doi:10.1039/b206642d · Cited by 19 · see citing works
Self-citationLee, A.-R. C. (2019) · Journal of Pharmaceutical Investigation · doi:10.1007/s40005-019-00438-y · Cited by 19 · see citing works
Kotuniak, R., Fra̧czyk, T., Skrobecki, P., Płonka, D., & Bal, W. (2018) · Inorganic Chemistry · doi:10.1021/acs.inorgchem.8b02841 · Cited by 15 · see citing works
Formation equilibria of nickel complexes with glycyl-histidyl-lysine and two synthetic analogues
Conato, C., Kozłowski, H., Świa̧tek-Kozłowska, J., Młynarz, P., Remelli, M., & Silvestri, S. (2003) · Journal of Inorganic Biochemistry · doi:10.1016/j.jinorgbio.2003.09.010 · Cited by 16 · see citing works
Self-citationJones, C. E., Zandawala, M., Semmens, D. C., Anderson, S., Hanson, G., Janies, D., & Elphick, M. (2016) · Biochimica et Biophysica Acta · doi:10.1016/j.bbagen.2015.10.008 · Cited by 12
“LHSALPF-NH2) has a histidine in the same position as the tripeptide Gly-His-Lys (GHK), a well-studied naturally occurring human copperbinding growth factor [28].”
Kukowska, M., Kukowska-Kaszuba, M., & Dzierzbicka, K. (2015) · Bioorganic & Medicinal Chemistry Letters · doi:10.1016/j.bmcl.2014.12.029 · Cited by 9 · see citing works
Synthesis and aggregation properties of a novel enzymatically resistant nucleoamino acid
Roviello, G., Mottola, A., Musumeci, D., Bucci, E., & Pedone, C. (2012) · Amino Acids · doi:10.1007/s00726-012-1219-9 · Cited by 7
“…A. Mottola D. Musumeci E. M. Bucci C. Pedone Istituto di Biostrutture e Bioimmagini, CNR, Via Mezzocannone 16, 80134 Naples, Italy e-mail: giovanni.roviello@cnr.it of enzymatically stable compounds useful for biomedical applications (Blankley et al. 1991; Braibanti et al. 1973; Conato et al. 2001).”
BackgroundBioactive oligopeptides and the application in skin regeneration and rejuvenation
He, Q., Liao, Y., Wu, Y., Zhang, H., Xiaohui, L., & Zhang, Y. (2025) · Journal of Applied Biomaterials & Functional Materials · doi:10.1177/22808000251330974 · Cited by 1 · see citing works
“73 After substituting 2-thiohis for His in GHK, the G-(2-thiohis)-”
Sialorphin and its analog as ligands for copper(II) ions
Kamysz, E., Kotynia, A., Czyżnikowska, Ż., Jaremko, M., Jaremko, Ł., Nowakowski, M. E., & Brasuń, J. (2013) · Polyhedron · doi:10.1016/j.poly.2012.10.055 · Cited by 4 · see citing works
Uğurlu, T., Türkoğlu, M., & Özaydın, T. (2011) · Drug Development and Industrial Pharmacy · doi:10.3109/03639045.2011.569934 · Cited by 3 · see citing works
Николаевна, Н. И., & Rizzarelli, E. (2025) · Antioxidants · doi:10.3390/antiox14121512 · Cited by 1 · see citing works
“However, GHK is characterized by a low survival capacity in human serum, making its delivery to target sites difficult, while GHK-Cu undergoes a rapid degradation in a weakly acidic environment or peptidase attack [223,224].”
Marciniak, A., Cebrat, M., Czyżnikowska, Ż., & Brasuń, J. (2012) · Journal of Inorganic Biochemistry · doi:10.1016/j.jinorgbio.2012.08.023 · Cited by 4 · see citing works
Formation and spectral properties of metal ion complexes of tripeptides
Hammouda, A. N., Jackson, G. E., Bonomo, R. P., & Elmagbari, F. M. (2016) · Inorganica Chimica Acta · doi:10.1016/j.ica.2016.07.043 · Cited by 2 · see citing works
Anxiolytic Effect of GHK-PGP Peptide and Its Structural Analogues in a Test with Punished Behavior
Alferova, M., Bobyntsev, I., Vorvul, A. O., & Tenkova, A. (2025) · Bulletin of experimental biology and medicine · doi:10.1007/s10517-026-06528-w · Cited by 1 · see citing works
Chelating Surfaces for Oriented Human Serum Albumin Molecules
Tuccitto, N., Messina, G. M. L., Destri, G. L., Wietecka, A., & Marletta, G. (2019) · Langmuir · doi:10.1021/acs.langmuir.9b00068 · Cited by 3 · see citing works
Mateescu, D.-M., Gavrilescu, D.-M., Mincioaga, R.-I., Pah, A.-M., Toma, A.-O., Serban, D.-V., Avram, A. C., Craciun, M.-L., Enache, B., & Mureșan, C.-O. (2026) · Pharmaceutics · doi:10.3390/pharmaceutics18091077
“Potentiometry, calorimetry, UV-Vis, circular dichroism, and EPR confirmed multiple stoichiometries and pH-dependent equilibria, with the lysine side-chain amino group contributing mainly at alkaline pH rather than under physiological conditions [16].”
Szczerba, P., Stefaniak, E., Płonka, D., Wezynfeld, N. E., & Bal, W. (2026) · Journal of Inorganic Biochemistry · doi:10.1016/j.jinorgbio.2026.113387
Peptides as unique tools to modulate redox levels: towards applications in living cells and in vivo
Balbali, N., & Delsuc, N. (2026) · Redox Biochemistry and Chemistry · doi:10.1016/j.rbc.2026.100079
Mishra, S., Sandey, L., Verma, A., Sahu, H., Rathor, S., Gajbe, B., & Prasad, J. (2023) · Journal of Chemical Health Risks · doi:10.53555/jchr.v13i6.2289
“[ 12] 2.”
- Authors
- Conato, C., Gavioli, R., Guerrini, R., Kozłowski, H., Młynarz, P., Pasti, C., Pulidori, F., & Remelli, M.
- Affiliations
- University of Ferrara (IT)
- University of Wrocław (PL)
- Corresponding author
- Remelli, Maurizio
- Venue
- Biochimica et Biophysica Acta (BBA) - General Subjects
- Publisher
- Elsevier BV
- Published
- 2001-05
- Volume
- 1526
- Issue
- 2
- Pages
- 199-210
- ISSN
- 0304-4165 (ISSN-L)
- OpenAlex
- W2079263573
- Cited by
- 55
- References
- 59
- Citations by year
- 2026: 4 · 2025: 4 · 2023: 2 · 2022: 2 · 2021: 1 · 2020: 4 · 2019: 2 · 2018: 3 · 2017: 2 · 2016: 2 · 2015: 1 · 2014: 3 · 2013: 1 · 2012: 2
- FWCI
- 0.89
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Biochimica et Biophysica Acta (BBA) - General Subjects (publishedVersion)
- PubMed (publishedVersion)
- Institutional Research Information System University of Ferrara (University of Ferrara) (submittedVersion)
- Topics
- Health Sciences › Medicine › Oncology › Metal complexes synthesis and properties
- Health Sciences › Nursing › Nutrition and Dietetics › Trace Elements in Health
- Physical Sciences › Chemistry › Organic Chemistry › Inorganic and Organometallic Chemistry
- Keywords
- copper complexes, tetrahydroisoquinolines, synthetic amino acids, complex formation equilibria, serum stability, growth factor activity
- MeSH terms
- Circular Dichroism, Copper, Growth Substances, Hydrogen-Ion Concentration, Oligopeptides, Peptide Hydrolases, Peptides, Protons, Spectrophotometry, Temperature, Thermodynamics, Drug Design
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
