The amount of money which California-based Sequoia Capital has invested has not been disclosed, but the deal is said to put a valuation of almost £500m on the Edinburgh company.
Even if Sequoia has only acquired a few percent of the existing share capital its investment is likely to run into tens of millions of dollars.
It is understood Skyscanner's founders - Gareth Williams, Bonamy Grimes and Barry Smith - all continue to hold shares.
Scottish Equity Partners (SEP) remains Skyscanner's single largest shareholder, six years after its initial £2.5m investment.
The most recent annual return filed at Companies House for Skyscanner Holdings shows SEP has around 44% of the ordinary shares, with Mr Williams at around 16% and Mr Grimes and his family at the same level. Mr Smith has around 6%.
Shane Corstorphine, Skyscanner's chief financial officer, said no new shares had been issued as part of the Sequoia deal but would not confirm which shareholders had offloaded parts of their stake.
Talks between both sides started two months ago and Mr Corstorphine characterised the deal as one of the biggest investments Sequoia has ever made.
He said: "We saw the opportunity of having a partner like Sequoia and the expertise that comes with that.
"Sequoia doesn't invest if they only think they will get [two or three] times the investment back.
"They are a long term investor - they believe they can make serious multiples. They are selective and we are immensely lucky they have picked us as a partner."
Mr Corstorphine said the business is on course to double revenue from the £33.5m recorded in 2012, with both the Asia Pacific and Europe zones performing strongly and a recently opened Miami office helping the company address the north and south American markets better.
But he also moved to quell suggestions the partnership with Sequoia would lead to Skyscanner looking for a stock market listing.
He said: "There are no plans to [do an initial public offering].
"We are focused on material growth and carving our own path."
Sequoia chairman, Sir Michael Moritz, who has previously served as a director at Yahoo and Google, is joining Skyscanner's board.
He said: "Skyscanner is one of the best technology companies ever to come out of Europe and is already a leading global player.
"Our job is to help take its current success a step further. Founders and CEOs like Skyscanner's Gareth Williams are part of an extremely rare species with talents that most of us yearn for but don't possess."
Sequoia, founded by Silicon Valley venture capital veteran Don Valentine in 1972, has previously invested in Oracle, Yahoo!, Cisco Systems, Electronic Arts, PayPal, YouTube, LinkedIn and Instagram.
Mr Williams said: "Sequoia Capital's investment in Skyscanner brings us a depth of experience drawn from a pantheon of tech brands."
Skyscanner has recently moved past the 300 mark in terms of employees as part of a plan to reach 500 over the next year.
A Glasgow office, with capacity for 120 staff, is to open in the next few weeks. As well as its Edinburgh HQ, it has the site in Miami plus operations in Barcelona, Singapore and Beijing.
Calum Paterson, managing partner at SEP and also a member of the Skyscanner board, said: "We have worked closely with Skyscanner since we first invested almost six years ago.
"During that time it has achieved exponential growth and been transformed into a world-class company
"We are pleased to welcome Sequoia Capital and Michael Moritz and we look forward to their contribution as we look to help Skyscanner achieve even greater success."
Skyscanner was founded in 2003 as a flights search engine and has since grown to include hotels and car hire.It acquired the Spanish-based hotel search specialist Fogg last month.