The 29-year-old sat level with Wayne Rooney on 53 strikes before England travelled to the Stadio Diego Armando Maradona for their opening Euro 2024 qualifier with Italy - renewing hostilities with the reigning champions nearly two years on from the Euro 2020 final.
Kane had a hand in the Three Lions' first goal on the night, as his blocked effort from a corner fell kindly for Declan Rice to fire home, and later in the first half, the Tottenham Hotspur talisman had prepared to meet Bukayo Saka's set-piece to the back post before Giovanni Di Lorenzo's arm got in the way.
Kane led the protestations to referee Srdjan Jovanovic, who awarded the spot kick after a look at the pitchside monitor, thereby presenting the striker with a golden opportunity to etch his name into English football folklore.
Kane - who became Tottenham Hotspur's all-time top goalscorer just last month - calmly sent Gianluigi Donnarumma the wrong way from the penalty spot to leave Rooney in his wake, scoring a record-breaking 54th England goal and letting emotion take over during the celebrations.
Speaking to Channel 4 after the game, the 29-year-old paid tribute to his "special" colleague Rooney while hailing his "magical moment" as he officially became the greatest goalscorer England has ever seen.
"It means everything. I was so excited to put the England shirt on and get back out here and get the campaign started for next year's Euros," the Tottenham striker said.
"It had to be a penalty and once it hit the back of the net, huge emotion. A magical moment. I was on the pitch when Wayne broke the record. I know what it meant to him and I was so proud of him.
"I remember when I gave him the boot for breaking the record he said he would be giving it back to me one day. A special man."
Despite entering the break with a comfortable 2-0 lead, England faced a barrage of Azzurri attacks in the second 45, and the hosts deservedly cut the deficit in half through debutant Mateo Retegui in the 56th minute.
Gareth Southgate's men were forced to see out the final 10 minutes with 10 men following Luke Shaw's sending off for two bookable offences, but the Three Lions ultimately got the job done to glean a slice of revenge for the Euro 2020 final.
Following the disappointment of a World Cup quarter-final exit, Kane believes that Thursday's win demonstrates that England are ready to go toe-to-toe with the big boys, adding: "Just a great night, we haven't won in Italy for so long and to score and win, so special.
"The World Cup was a difficult way to end but we spoke this week about getting back to it. We are one of the best teams in Europe. Getting this win shows we are ready for the challenge."
Meanwhile, manager Southgate affirmed that he "could not be happier" for his record-breaking captain, saying: "To break the record in the manner he did given the recent history is an indication of his strength of character and mentality. I couldn't be happier for him and his family, he is a brilliant professional."
England sit top of the Group C rankings thanks to their opening win, and Southgate's men now prepare to welcome Ukraine to Wembley on Sunday evening.