Skin Proteins Reveal How Mummies Died

Proteins from the mummies' skin and muscle samples show the people likely had cancer, lung infections and other diseases.

An international team of researchers has identified hundreds of proteins in skin and muscle samples from 4,200-year-old Egyptian mummies, finding signs of diseases that may have caused their death.

Published in the journal Philosophical Transaction of the Royal Society A, the groundbreaking study shows that proteins isolated from ancient mummified tissue can reveal inflammation, immune response and possibly cancer.

The researchers collected four skin samples and one muscle biopsy from three mummies stored in the Egyptian Museum in Turin, Italy.

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Dating back to the First Intermediate period (about 2181–2055 B.C.), the mummies were excavated in cemeteries at Assiut and Gebelein between 1911 and 1920 by an Italian archaeological mission led by Ernesto Schiaparelli.

The Assiut mummies, a female known as Khepeshet and a male known as Idi, came from elite burials and were interred, with grave goods, in sealed and decorated wooden coffins.

In contrast, the mummy from Gebelein, an unknown adult individual, was buried in a coffin made out of a hollowed out tree trunk.

"All these mummies are in poor condition, but that is what made them perfect for retrieving biopsies without causing further damage," Jana Jones, from the Department of Ancient History at Macquarie University, Australia, told Discovery News.

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Analysis showed that all five samples contained large numbers of collagens and keratins, confirming previous studies that identified these proteins as very long-lived.

Overall, the researchers identified more than 230 proteins in the 4,200-year-old samples, finding evidence for inflammation, infection and possible cancer.

Jones and colleagues Paul Haynes and others from the Department of Chemistry and Biomolecular Sciences, Macquarie University, Raffaella Bianucci, at the Legal Medicine Section of the University of Turin, Italy and Dong Hoon Shin, at the National University College of Medicine in Seoul, South Korea, estimate that any proteins observed at higher abundance in mummified samples of that age must have been expressed at relatively high levels in the original tissue.

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"Using that approach, we have been able to show that many of the proteins still present in these samples are linked to inflammation and immune response," the researchers wrote.

Analysis of skin tissue from the mummy known as Khepeshet identified a protein signature indicative of a severe immune response.

"A subset of those proteins were strongly linked to bacterial infection in the lungs," Paul Haynes said.

He noted there is a strong possibility that Khepeshet was suffering from a bacterial pulmonary infection, such as tuberculosis.

"This is something you could point to as a possible cause of death," Haynes said.

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Most likely, the mummy known as Idi was also suffering from a life-threatening disease.

Analysis of both skin and muscle samples identified numerous proteins associated with inflammation and severe immune response.

In the muscle sample in particular, the researchers found two proteins, DMBT-1, which functions as a tumor suppressor, and transglutaminase.

Haynes explained that increased abundance of both DMBT-1 and transglutaminase is generally correlated with pancreatic cancer progression.

"This allows us to speculate that Idi may also have been suffering from pancreatic, or some other cancer," Haynes said.

Few proteins were identified for the third mummy, so the researchers were unable to find details about the cause of death.

"The remains were interred in a hollowed out log rather than a sealed coffin. The mummy would have been exposed to the elements over time and this may have caused protein degradation," Jones said.

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She noted the First Intermediate period was Egypt's first "Dark Age."

"It was marked by political unrest, changed economic conditions, mega drought and famine," Jones said.

Although little is known about the health of the population in this period, it is no mystery that food and water shortages weaken the immune system, paving the way to infectious diseases such as malaria, tuberculosis, visceral leishmaniasis and other parasitic intestinal infections.

Groups affected by these chronic conditions are at increased risk of contracting cholera, typhoid fever and acute respiratory infections.

"Our study provides a historical context for medical conditions that are still found in the modern world," Jones said.

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Discovered in 1881 in the Deir el-Bahri Cache above the Mortuary Temple of Hatshepsut and unwrapped by Gaston Maspero in 1886, this mummy, known as CG 61066, was badly damaged by grave robbers in antiquity. The left arm was broken off at the shoulder, the right arm cut off at the elbow and the right leg severed from the body.

The mummy's name remains uncertain. It was identified as that of King Thutmosis II (reign c. 1493–1479BC) thanks to a wrongly spelled label. However, the inscription appear to have overwritten an earlier sign referring to Thutmosis I. This could indicate the inscription had been changed from Thutmosis I to Thutmosis II, suggesting that the royal mummy CG 61066 is that of Thutmosis I rather than his likely son Thutmosis II.

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Badly damaged by ancient robbers, this mummy had fallen into pieces, the well preserved head broken off, all four limbs detached and the feet severed. The mummy has been identified as that of Thutmosis III, the sixth pharaoh of the Eighteenth Dynasty (reign about 1479–1425 BC) and shares a striking resemblance with Thutmosis II (mummy CG 61066), his purported father.

The identification is based on the fact that when it was unearthed, the mummy was lying in a coffin bearing in its interior traces of inscriptions made for Thutmosis III. A linen shroud with a funerary book which certainly belonged to Thutmosis III, was possibly placed there by the embalmers.

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French excavator Victor Loret found the mummy CG61069 in 1898 in the KV35 tomb in the Valley of the Kings. The mummy was lying in a wooden coffin placed into a stone sarcophagus with inscriptions naming Amenhotep II, the seventh pharaoh of the 18th dynasty of Egypt, who ruled between 1428 and 1397BC. A simple label on the mummy's shroud presented the identity of Amenhotep II.

Some doubt however remains. The coffin did not give a name and was too large for the mummy. Moreover, the faded hieratic ink inscription could have been easily misread. "With reservations the mummy CG 61069 should be considered as Amenhotep II until proven otherwise," Ruhli and colleagues concluded.

Found in 1898 in the KV 35 tomb, this damaged mummy -- both feet are broken off and the right leg was ripped off at the knee joint -- shows the face of an extremely emaciated man. The body has been identified as Thutmosis IV, the 8th Pharaoh of the 18th dynasty who ruled between 1397 and 1387 BC, thanks to inscriptions on the chest and coffin correctly spelled and clearly visible. According to the researchers, the identification should be considered reliable.

When it was unwrapped in 1905, this mummy revealed a body in rather bad condition. The head was broken off, most of the soft tissue from the face gone, the right leg was cut away from the trunk, and part of the foot missing. The embalmers taped the mummy together in the attempt to restore a lifelike appearance. Bird bones, a human big toe, and parts of an arm were found inside the body cavity. The genetic profiling determined the mummy was the consort of Queen Tjye and as the genetic father of the mummy from tomb KV 55, thus identifying the body as Amenhotep III.

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The identity of these exceptionally well-preserved mummies is certain, since the coffins and funerary objects bear their names and status. Genetic testing recognized Yuya and Thuya as King Tut's great-grandparents.

This nameless and naked mummy was found in 1898 in the cachette KV 35 together with the so-called Younger Lady and a teenage boy. The regal quality of mummification and her bent arm recognized her as a queen; speculations were made to identify her as of Tjye, Nefertiti, or Hatshepsut. In view of the genetic test, where she was proved to be the daughter of Yuya and Thuya, the identification as Tjye, wife of King Amenhotep III, appears the most realistic one.

This body, consisting of a completely disarticulated skeleton with few missing part and a damaged skull, was identified as Akhenaton although some argued he could have been the elusive pharaoh Smenkhkare. In the Tutankhamun Family Project he was proved with molecular genetics to be a direct descendant of the "Elder Lady" (Queen Tiye) and mummy CG 61074, commonly regarded as "Amenhotep III," and this would indicate that he was the heretic king Akhenaton.

The KV 55 mummy was also determined to be the genetic father of Tutankhamun. Inscriptions support the genetics. Not only the body found in KV55 bore golden bands with the name Akhenaton, but inscriptions from Tell el-Amarna, the city of the heretic king, describe King Tut as the son of Akhenaton.

Found by Victor Loret in 1898 in tomb KV35, the mummy was considered to be an unknown royal family member of Amenhotep II, because she was found in his burial. In 1999, Marianne Luban suggested the mummy might be Queen Nefertiti, based on her profile resemblance with the famous Berlin bust. In 2004 Egyptologist Joann Fletcher also proposed the mummy is Nefertiti, relying her theory on portable x-ray, forensic face reconstruction and resemblance between art and mummy. Her identification raised much controversy.

The DNA analysis revealed the mummy is the mother of Tutankhamun, but did not offer an identity. In their meta-analysis, Rühli and colleagues also present her as Queen Nefertiti. "We can't be fully certain of her identity, however inscriptional evidence and facial resemblance with Tutankhamun as seen in CT scans, strongly suggests the mummy belongs to Nefertiti," Rühli said. "Nefertiti is labelled in inscriptions to be Tutankhamun's mother and indeed the mummy known as the Younger Lady is genetically suggested to be King Tut's mother," he added.